- Email: [email protected]
Functional domains of cell adhesion molecules
Functional
domains
Yoshihiko National
of cell adhesion
Yamada Institutes
and Hynda
of Health,
Bethesda,
molecules
K. Kleinman Maryland,
USA
A number of molecules involved in cell adhesion (e.g. fibronectin, laminin, collagens I and IV, thrombospondin, entactin) have now been identified and the consequent roles that they play in the processes of growth, migration, differentiation and tumor spread have been described. Active sequences of the molecules have been identified using synthetic peptides derived from specific domains. Several adhesive molecules contain multiple active domains with different biological activities.
Current
Opinion
in Cell
Biology
Introduction
Laminin
Recently, synaptic (s-laminin) and merosin, homologues of the Bl and A chains, respectively, have been described in various tissues [ 16-181. S-laminin contains an L.RE sequence that promotes the adhesion of ciliary ganglia [19,20].
Iaminin is a large glycoprotein (molecular weight = SOOOOO),which is found primarily in basement membrane, the thin extracellular matrix that underlies epithelial cells and surrounds muscle, fat and peripheral neuronal cells. Iaminin is therefore in contact with a variety of cells and it is not surprising that it contains multiple active sites [ 1,2]. A major form of laminin is composed of three chains designated A (molecular weight = 400000), Bl (molecular weight = 210000) and B2 (molecular weight = 200000). Laminin has various biological activities including the promotion of cell adhesion, migration, growth, neurite outgrowth, collagenase N activity, tyrosine hydroxylase activity and tumor metastases. Several of the active peptide sequences are Current
Biology
4:819823
distinct in their cellular response and the type of cell with which they interact. For example, the Bl chain peptide, YIGSR, does not interact with neuronal cells, whereas the A chain peptide, PA22-2, which contains SIKVAV, and the B2 chain peptide p20 recognize neuronal cells [ 1,3,4]. Likewise YIGSR, PDSGR and F9 (RYWLPR) are located on the Bl chain and inhibit tumor metastases [1,2,5,6,7-J], whereas PA22-2 increases tumor metastases, growth, angiogenesis, plasminogen activation and collagenase N activity [ 9,10,11**]. As YIGSR and SIKVAV have opposing activities in tumor metastases and angiogenesis, it is likely that the two sites are not available to cells in tissues at the same time, but rather are developmentally regulated by the presence of other matrix components. It is interesting to note that the all-D-con&tired IKVAV-containing peptide has similar biological activity to the al-L-IKVAV peptide [ 121. Various active peptides have been defined from the carboxyl globular domain of the A chain (termed G domain) and all appear to be active with neural cells [ 12,13*,14]. It should be noted that this domain has significant homology with other proteins including S protein, androgen-binding protein and sex hormone-binding globulin [ 15.1. An additional point to keep in mind is that some cells, such as endothelial cells, can recognize multiple sites on laminin including YIGSR, RGD and SIKVAV ( [ 16); HK Kleinman et al., unpublished data).
The extensive study of several adhesive macromolecules has been prompted by their widespread involvement in development and diseases. This review will describe the more abundant and better studied extracellular matrix components laminin, Iibronectin, collagens 1 and N, thrombospondin and entactin/nidogen, and their active sites defined to date by short synthetic peptides. Emphasis will be placed on the responding cells because active sites vary in cell types with different specificity. All of these components have been sequenced, which has facilitated the approach of employing synthetic peptides. Part of this review will focus on one of the laminin-derived synthetic peptides containing YIGSR (where YI is tyrile and GSR is from the single-letter amino acid code), which is more active in the polymeric form and has been found to block the differentiation of some cells, tumor growth and angiogenesis. Iaminin, tibronectin and the collagens contain multiple biologically active sites with distinct activities and cell specificities. In some cases, the cellular receptors for a particular site differ between specific cells, which leads to a large diversity of biological functions and cell regulation.
@
1992,
The MGSR site on the laminin Bl chain has been studied by several laboratories because of its potent inhibitory effect on tumor metastases [5,6,7*] and growth 1211. This peptide blocks tumor metastases by reducing tumor cell adhesion to laminin [4,22], but inhibits angiogenesis [7*,17,23]. The activity of this peptide has been increased by preparing it in the cyclic form, as a polymer, coupled to polyethylene glycol and as a ‘starburst’ structure [6,24,25]. It is not clear if the increased activity is due to a better conformation, a longer half-life Ltd
ISSN
0955-0674
819
820
Cell-to-cell
contact
and extracellular
matrix
. rable
1. Cell
attachment
tiolecule
sequences
in extracellular
matrix
Recognition (single-letter
-aminin
proteins. Cell
sequence amino
acid
type
distribution/function
code
Bl
YIGSR
Many
YICSR
cells,
RYVVLPR
F9
PDCSR
Synergistically
LCTIPC
Binds
PDGSR LCTIPC
site
inhibits
Heparin
metastasis
binding
functions
with
to eleastin
YICSR?
receptor
Laminin 82 RNIAEIIKDA
P20
Promotes
neurite
Ciliary
ganglion
outgrowth
Laminin s LRE site Laminin
LRE
neurons
A
PA22-2
IKVAV
Promotes
neurite
outgrowth,
activation RCD
site
Endothelial
RGDN
GD-1
KATPMLKMRTSFHCCIK
CD-2
KEGYKVRLDLNITLEFRUSK
CD-3
KNLEISRSTFDLLRNSYCVRK
CD-4
DGKWHTVKTEYIKRKAF
CD-6
KQNCLSSRASFRCCVRNLRLSR
and
Promote
plasminogen
tumor cells
neurite
formatlon and
others
outgrowth,
heparin
blnding i Melanoma
cells
Fibronectin RGD
site
RGDS
CSI
site
LDV
REDV
site
Synergestic
Most Most
cells,
neural
REDV site
Not
yet
inhibits
metastasis
crest-derived
cells,
Melanoma Most
determined
cells
Peptide
I
YEKPGSPPREVVPRPRPGV
Melanoma
cells
Peptide
II
KNNQKSEPLIGRKKT
Melanoma
cells
Collagen
IV
IV-H1 Hep
GVKGDKGNPGWPGAP Ill
Melanoma
GEFYFDLRLKGDK
cells,
promotes
Heparin
binding
Collagen
I
RGD
site
RGDTP
Fibroblasts
and
RGD
site
SRGRTG
Fibroblasts
and
DGEA
lymphocytes
cells
site
DGEA
Platelets
and
motility
others others others
Thorombospondin VTXG RGD
site site
WSXW Entactin RGD
VTXG
Promote
platelet
aggregation,
metastasis
RGD site
wsxw
Heparin
binding
(Nidogen) site
FRGDS
in the bloodstream, or a combination of both. Some difficulties have been encountered with the synthesis and stability of the pentameric molecule and this has led to some inconsistencies for its activity. The YIGSR peptide has also been found to interact with normal cells and to influence cell differentiation. As already mentioned, angiogenesis is blocked in zUt!o in the chick chorio-allantoic membrane and in a rabbit eye model; in addition, a specific stage of sea urchin embryonic development, archenteron elongation, is blocked while other stages are unaffected [ 70,261. The migration of endocardial cushion mesenchyme and chick neural crest from the notochord is also blocked by YIGSR
Mammary
tumor
cells
[27,28]. Many cells are observed to differentiate when plated on a Iaminin-rich basement membrane matrix [29]. In some, but not all, cases, YIGSR can block this differentiation. For example, Sertoli cell cord formation is blocked by MGSR due to a decreased cell attachment to the matrix [30]. Similar results are observed with bone cells [ 311, whereas endothelial cells remain attached to the matrix but do not align [ 161. In contrast, the formation of salivary gland-like structures on basement membrane is unaffected by MGSR, but is blocked by the SIKVAV-containing peptide [ 32.1. These &ta demonstrate that YIGSR is one of the major functional sites in laminin for a variety of cells.
Functional
Table 2. Activities
of laminin-derived
YIGSR peptide.
Activity
Cell type
Promotes
adhesion
HT1080,
or system
CHO,
endothelial
cells.
trophoblasts Blocks
laminin
adhesion
OVCAR-3,
colon
cell lines, PMN,
macrophages Promotes
migration
B16-FIO.
chick
endocardial
cushion
mesenchyme Inhibits
tumor
growth
lnhiblts
angiogenesis
B16-FlO,
Tsu-prl,
small
carcinoma, Chick
sarcoma
chorio-allantoic rabbit
cord
formation
Inhibits
urchin
Inhibits
outgrowth
development
vein
Primary Sea urchin
180 membrane,
eye model,
unbilical Inhibits
cell lung,
adhesion
molecules
Yamada and Kieinman
peptide blocks platelet adhesion to collagen, but not to fibronectin or laminin. Surprisingly, it also blocked T47D breast adenocarcinoma cell adhesion to collagen I and laminin. Both platelets and the T47D cells use the same receptor (azpI integrin) in their interaction with this sequence. Thus, the ability of cells to interact with this site is dependent on the presence of a specific receptor. The DGEA sequence recognizes the azpl integrin. This sequence is not in laminin, which suggests that a2pI can recognize additional sequences. Collagen N also has several active sites [2,42]. A major cell-binding site interacts with both the alpI integrin and the al& receptor [42,43**]. The specificity among various cell types has not yet been determined.
cells archenteron
elongation trunk
neural
crest
Fibronectin Fibronectin (molecular weight = 540000) is a serum protein that is also found in fibrous connective tissue and in the basement membrane of certain tissues [ 1,331. Like laminin, it has multiple active sites with different cellular specificities. The RGD sequence in libronectin was the first sequence to be reported active in promoting cell adhesion [34]. This sequence is found in more than 100 other proteins, including laminin, collagens I and N. entactin, vitronectin and fibrinogen [ 11. As some of these proteins are not biologically active, it would appear that the sequences and structure surrounding RGD are important for the activity in the native molecules. In fact, several upstream sequences in fibronectin have been reported to function synergistically with RGD in tibronectin [35-l. Additional active sites on fibronectin have been described, including LDV, which are cell-specific and in some cases located in alternatively spliced domains 136,371. All of the active sequences identified in libronectin have so far been found mainly to promote cell adhesion and sprcJding. The RGD site can reduce tumor metastases [ 38,391, but it is less active than the laminin-derived YIGSR.
Collagen
of cell
cell in vitro
Sertoli
embryo
Stage 12 chick
domains
I and IV
Collagens have been used to coat culture dishes because they promote cell adhesion and survival. Collagen I forms a Iibrillar network, often with fibronectin in L@ljo,whereas collagen N forms a complex non-Iibrillar network. Besides the RGD sites that are present on both collagens [2,-iO], type I collagen has a DGEA site, which is active with platelets and certain other cells [41**]. This
Thrombospondin
and entactin
(nidogen)
Thrombospondin is a large glycoprotein (molecular weight = 450 000) that is released from platelets. It is also present in various extracellular matrices due to its interactions with heparin, fibronectin and collagen. It has been found to promote cell adhesion and various other biological events [44,45-,46-l, including neurite outgrowth and inhibition of tumor metastases. To date, three cellbinding sites have been defined, and two of these are in the carboxyl-terminal heparin-binding domain. One peptide that is constructed from the type I repeat WSXW promotes melanoma cell adhesion, whereas another type I repeat peptide CSVTCG and its homologues both promote adhesion and inhibit platelet aggregation and tumor metastases [ 46.0,471. Entactin/nidogen (molecular weight = 158000) is found in all basement membranes, where it binds to other bioactive components including laminin and collagen N. It also binds well to collagen I. The RGD site on entactin is functional with various cells [43**,48-501. Due to its presence in complexes with other bioactive matrix molecules, it can be expected that some synergy in activity exists although that has not yet been demonstrated. The interaction of enactin with tibronogen chains also suggests a role in hemostasis [49].
Conclusions A number of cell adhesion molecules are multifunctional and demonstrate cell type specificity. Several active sites have been identified by synthetic peptides and others probably exist. The definition of active sites and duplication of their activities by small synthetic peptides has helped to unravel specific developmental mechanisms. Such peptides may also be used clinically in the treatment of diseases, clotting problems, or to coat prostheses and implants for better tissue adherence. These peptides may also be used to improve wound healing and/or reduce scarring.
821
822
Cell-to-cell
contact
References
and extracellular
and recommended
Papers of particular interest, review, have been highlighted . of special interest .. of outstanding interest 1.
matrix
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reading within
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1-l.
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3.
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4.
;finin tide.
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.I 13iol Chew
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integrin
ajpl
was found
to bind
to the pep-
15. .
Jos~:.I+I DR, BAKER ME: Sex Hormone Binding Globulin, Androgen Binding Protein and Vitamin K-dependent Protein S Are Homologous to Laminin A, Merosin. and Drosophila Crumbs Proteins. /+l.Sf?B ./ 1992. 6:21?7-2481. Strong homologies in the G domain (carboxTl terminus) of the A chain of Iaminin are obsencul nith various proteins. 16
GKANT
DS. T&+IIRO K. Secili-REAL B. YM~ADA I’, MAK~IN GR. I1K: Two Different Laminin Domains Mediate the Differentiation of Human Endothelial Cells into Capilliarylike Structures. Cc/I 1989, 58:933-943.
tiINhL.w
17.
SANES J, ENG\‘AU E, BLI~OWWI erogeneity of Basal Laminae: lagen IV at the Neuromuscular Cell Bid 1990, 111:1685-1699
18.
HL’NTER DD, SHAH V, MERUE JP. SANES JR: A Laminin-like Adhesive Protein Concentrated in the Synaptic Cleft of the Neuromuscular Junction. Nn!lrrr 1989, 338:223-23-i.
Effect of on the In1 ./ Rio/
19.
HIIN~L!R DD. POR-IZR BE, B~II~K JM. ADUIS SP. MER~E JP, SAIIES JR: Primary Sequence of a Motor Neuron Selective Adhesive Site in the Synaptic Basal Lamina Protein S-laminin. Cell 1989, 59905-913.
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20.
HUSKER DD, C~HXIAN N. MORRIS-V,UERO R, Bti~.oclc JW. AD,u.~s SP. SANES JR: An LRE (Leucine-Arginine-Glutamate)-dependent Mechanism for Adhesion of Neurons to S-laminin. J Newosci 1‘991, 12:396&3971.
5.
IWAMOTO
Y, MARTM of Iaminin 328:1132-l
6.
Y, Ro~t7’ FA, GR~ J. SUAKI M. KIJZIN~IAN HK, Y&\IAI)A GR: YIGSR a Pentapeptide from the Bl Chain Inhibits Tumor Cell Metastases. Science 1987, 134.
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7.
.
8.
SKlrBrrz APN, ~McCAR~~~ JB, ZHOA Q. YI X. FIIRCHT LT: Definition of a Sequence RYVVLPR, within Laminin F-9 that Mediates Metastatic Fibrosarcoma CeU Adhesion and Spreading. Cancer Res 1990, 50~7612-7622.
9.
KANEMOTO T, REICH R. GREA~OP&X D. AOLER SH. YA~u~,A Y. KLEINMAN HK: Identification of an Amino Acid Sequence from the Laminin A Chain which Stimulates Metastases Formation and CoUagenase IV Production. Proc Nor/ Au& Sci USA 1990, 87:227’+2283.
10.
STACK S, GRAY RD, PKZO SV: Modulation of Plasminogen Activation and Type IV CoUagenase Activity by a Synthetic Peptide Derived from the Laminin A Chain. Biochetnimy 1990, 30:2073-2077.
11.
K~OTA S, TA~HIRO K, YA&IADA Y: Signaling Site of Laminin with Mitogenic Activity. J Bid chetn 1992, 267342854288. ze laminin A chain peptide containing SIKVAV stimulates PC12 cell groti and mRNA expression of c-/a\- and c-&n. The binding of both the-se proto-oncogene products to the API site m also increased b) the peptide. 12.
NOMIZU M, UTANI A, SHIRA~SHI N, KIBBEY MC, YAMADA Y, ROUER PP: The AU-D-COntiguratiOn Segment Containing the IKVAV Sequence of Laminin A Chain has Similar Activities to the AU-L-peptide in Vitro and in Vivo. J Biol C!wn 1932. 267:1411&14121.
21.
FR~D~IAN R. G~ACONNE G, K..mh!OTO T. MARTIN GR. G,~DAR AF, MLIISHINE JL: Reconstituted Basement Membrane (Matrigel) and Laminin Can Enhance the Tumorogenicity and Drug Resistance of Small CeU Lung Cancer Lines. Proc N&l Acad Sci I’SA 1990, 87:669ti707. Describes a new method for growing tumor cells that employs subcutaneous co-injection of tumor cells and basement membrdne. The peptide IlGSR blocks the growth of these tumors when co.injected whereas the peptide MGGR is without effect. 22.
DANEKEK DW, Pwzw AJ. SMELL: GD, MERC~IRIO AM: Relationship between Extracellular Matrix Interactions and Degree of Differentiation of Human Colon Carcinoma CeU Lines. Cuncer Res 1989, 496814.
23.
SWENR’
GEHEEN KR. SIURAMARAo P, FURCHT LT, SKLIBI-I-~ APN: A Synthetic Peptide Derived from the Carboxyl Terminus of the Laminin A Chain Represents a Biding Site for a$, Integrin. J Cell Biol 1992, 117:44’+459. A peptide from the globular domain of the laminin A chain supponed cell attachment and blocked laminin-mediated cell adhesion; antibody to the peptide also blocked laminin-mediated adhesion. Using peptide
TM, KIBBEI’ MC, ZAIN M, FwDhtU‘I R. KLEIN~WN HK: Membrane and the Laminin Peptide Containing Promote Tumor Growth and Metastases. Cancer Meluskzsis Reel 1991, 10:245-254. Basement SIKVAV
23.
KAW~AKI K. NA~~AKAWA M, MUMI T. M~;~AITA H, IWAI Y. Hahn\ T, MA~~I~II T: Amino Acids and Peptides XIV, Laminin Related Peptides and Their Inhibitory Effect on Metastases Formation. Bicdem Biopqa Res Commun 1991, 174:115%1162.
25.
KwNhlAh’ IHK, GRAI; J, IWAMOT’O Y. SASAKI M, SCHASTEEN CS, YMLADA Y. MARnN GR, RO~EY FA: Identification of a Sec. ond Active Site in Lam&n for Promotion of CeU Adhesion, Migration, and Inhibition of in Viuo Melanoma Lung Colonization. ArdJ Biocbem Bicgdga 1989. 272:39-45.
26.
BENX)N S, CHLI~PA S: DUTerentiation of in Vitro Sea Urchin Micromeres on Extracellular Matrix in the Absence of Serum. J Eq Zwl 1990, 256:222-226.
27.
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28.
BILOZUR ME, HAY ED: Neural Crest Extracellular Matrix Utilizes Laminin, lagen. Deaf Biol 1988, 174:1159-1162.
13. .
K, HUNTER D: Molecular Hetlsoforms of Laminin and ColJunction and Elsewhere. J
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KwNhwi-4 HK, GRAY J, IWAMOTO Y. K~I-~EN GT. om RC, SASAKI M, YAMAHA Y, MARTIN GR, LIICIXNI~IU-EDDS L: Role of Basement Membrane in Cell Differentiation. N I’ Acud Sci 1987. 513:134-145.
30.
HADLEY MA, WEEKS BS, KLEINMAN HK, motes Tubule Formation by Cultured 1990, 140:31%327.
31.
V~IKICRIC C, LLJITEN FP. KIEIN~WN HK. REDDI tiation of CanaUcuU-like Bone Cell Network Membrane Matrix Components and Defined Laminin. Gel/ 1990. 63:437-i45.
DYM M: Laminin ProSertoU Cells. Deft Riol
AH: Differenby Basement Domains of
32.
KIBH~Y MC, ROYCE LS, DYhi M, BALIM BJ. Ku1Nhi.m HK: Glandular Morphogenesis of a Human Submandibular Cell Line by Basement Membrane Components in V&o. E@ Ceil Kes 1992, 198:343-357. A human submandibular cell line forms glandularlike structures on a basement membmne substrate. The laminin peptides MGSR and RGD have no effect on the morphology of the cells, whereas the SIKVAV pep tide blocks gland formation and causes a rapidly growing monolayer of cells to form. .
33.
PIERXHBACHER of Fibronectin merits of the
34.
YUIADA proteins. by Hay
KM: In E.D.
MD, RIIOSWHTI E: Cell Attachment Activity Can be Duplicated by Small Synthetic Frag Molecule. Nntfrre 1984. 309:30-33. Fibronectin Cell Bidon? New York:
and
Other Cell Interactive of fl,e I:v!rucell~rlcrr .Ilrr/ris Plenum Press; 1991:111-l-16.
GlycoEdited
AOTA S, NACAI T, YA~MDA KM: Characterization of Regions of Fibronectin besides the Arginine-Glycine-Aspartic Acid Sequence Required for Adhesive Function of the Cell-binding Domain Using Site-directed Mutagenesis. ./ Bid Cheer 1991, 266:1593%15943. Using deletion analysis and oligonucleotide site-directed mutagenesis, regions on libronectin were found to be required for full activity of the RGD site.
35.
..
36.
37.
HLlhIPHlUES MJ, kCIYAh!A SK, KOhlORlYA A, OlDEN K, \I’A!\lAI,A KM: Identification of an Alternatively Spliced Site in Human Plasma Fibronectin that Mediates CeU Type-specific Adhesion. J Cell Rio/ 1986. 103:2637-2(~7. HLIhwHms MJ. KO~IOR~‘A A, AK~‘~IA SK, OIIXIN K. Y.~\!AI)A KM: Identification of Two Distinct Regions of the Type 111 Connecting Segment of Human Plasma Fibronectin that Promote CeU Type-specific Adhesion. J Hi01 C/~em 1989,
domains
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40.
of the Peptides Murine
AGRU. MV, RATES RC. Bow AW, BARNS GF: Arg-Gly-Asp-containing Peptides Expose Novel Collagen Receptors on Fibroblasts Implications for Wound Healing. Cell Ke@rl 1991, 12:1035-1044.
adhesion
Yamada
molecules
ST’AATZ WD. Foli KF, ZULTER MM, SANr’oTo SA: Identification of a Sequence for the a,Pl Integrin 1991, 2667363-7367. Peptides preparcil from cyanogen bromide lagen identilird a four amino acid sequence and other cells in their a#, integrin-mediated also blocked T47D breast adenocarcinoma
?l. ..
42.
and Kleinman
ADA\IS SP, RODRIQIIU. BA, Tetrapeptide Recognition in Collagen. J No/ Cbem fmgment CB3 of type 1 col( DGEA) used by platelets adhesion, This peptide adhesion to laminin.
CHEIXRC MK, hlcC,%aT!n’ JB, SI(t!BrrL APN. FL~RCHT LT. TSILIHARS EC: Characterization of a Type IV Collagen that Promotes Melanoma CeU Adhesion, Spreading and Mobility. ./ CeN Rio1 1991. 111:201-270.
t3.
VANLIENBERC P. KEKN A, RIES A, LUCKENBIU.EDDS I+ WN K, K~IHN K: Characterization of a Type N Collagen Major CeU Binding Site with Affinity to the A, B, and the &PI lntegrins. J Cell Biol 1991, 113:1475-1483. A cyanogen bromide fragment CB3 of collagen IV is ils active as the intact molecule in promoting cell adhesion. Using CB3 affinity chromatogmphy. two integrins. alal and a&- - were isolated. ..
i-1.
Trlsn?ls~~ L. Shim-1 Promoted 236: 15X-
GP. ROlJlhtiN Vl., Mt’lU’~ A, SlEGlIR K. ShllTH S. KORIXWSKI J, L~LIDSEN KA: Thrombospondin CeU Substratum Adhesion. Science 1987, 1573.
45. ..
O’SHM KS, LLII L-H. DIN-~ VM: Thrombospondin and a 140 kD Fragment Promote Adhesion and Neurite Outgrowth from Embryonic Central and Peripheral Neurons and from PC12 Cells. Neurojr 1991, 7:231-237. Neurite outgrowth did not involve the RGD sequence or the aminoterminal heparin.binding domain.
-16.
TI’S~YNSKI GP. Ro7HhIAN VL, DELIXH AH, HA~IILI’ON BK, EYAL J: Biological Activities of Peptides and Peptide Analogues Derived from Common Sequences Present in Thrombospondin, Properdin, and Malarial Proteins. J Cell Bioll992, 1 16:209-2 17. Thrombospondin is produced by platelets and influences platelet aggre+on, cell adhesion and tumor metastases. Two active sites, which are also present in a number of other proteins including properdin and malarial proteins. contain the acthig. ..
-i7.
GL,O N. KHI’T~XH, NEC,II~: E, VOGEI. T. 1~~ D& ROBERIS DD: Heparin and Sulfatide-binding Peptides from the Type I Repeats of Human Thrombospondin Promote Melanoma CeU Adhesion. Proc N&l Auzrl Sci I’SA 1992. 89:304&3013.
ix.
OIAKR%\‘AHTI S. T~L\I MF. CHEIINC; AE: The Basement Membrane Glycoprotein Entactin Promotes CeU Attachment and Binds Calcium Ions. J Biol C/Jet,? 1990, 265:1059?-10603.
-19.
,~L!NN K, DEIVJIANN R. A~:DWI.IEY M. TNPL R, RNhIoml L, YMIADA Y. P&v TC. CohwAY D. CHI: ML: Amino Acid Sequence of Mouse Nidogen, a Multidomain Basement Membrane Protein with Binding Activity for Laminin. Collagen N and Cells. EVBO J 1989. 8:65-‘7.
50.
Wr’ CY. CHL:NG AE: Potential Role of Entactin in Hemostasisspecific Interactions of Entactin with Fibrinogen A-a and B-P Chains. ,/ Rio/ (3enr 1991, 266:18802-18807.
262:68866892. 38.
of cell
Y Yamdda and HK KleinnXIn. laboratory National Institute of Dental Rt!se’drch. Bethesda, Mar&and 20892, USA
of Developmental National Institutes
of
Biology, Health.
823
domains
Yoshihiko National
of cell adhesion
Yamada Institutes
and Hynda
of Health,
Bethesda,
molecules
K. Kleinman Maryland,
USA
A number of molecules involved in cell adhesion (e.g. fibronectin, laminin, collagens I and IV, thrombospondin, entactin) have now been identified and the consequent roles that they play in the processes of growth, migration, differentiation and tumor spread have been described. Active sequences of the molecules have been identified using synthetic peptides derived from specific domains. Several adhesive molecules contain multiple active domains with different biological activities.
Current
Opinion
in Cell
Biology
Introduction
Laminin
Recently, synaptic (s-laminin) and merosin, homologues of the Bl and A chains, respectively, have been described in various tissues [ 16-181. S-laminin contains an L.RE sequence that promotes the adhesion of ciliary ganglia [19,20].
Iaminin is a large glycoprotein (molecular weight = SOOOOO),which is found primarily in basement membrane, the thin extracellular matrix that underlies epithelial cells and surrounds muscle, fat and peripheral neuronal cells. Iaminin is therefore in contact with a variety of cells and it is not surprising that it contains multiple active sites [ 1,2]. A major form of laminin is composed of three chains designated A (molecular weight = 400000), Bl (molecular weight = 210000) and B2 (molecular weight = 200000). Laminin has various biological activities including the promotion of cell adhesion, migration, growth, neurite outgrowth, collagenase N activity, tyrosine hydroxylase activity and tumor metastases. Several of the active peptide sequences are Current
Biology
4:819823
distinct in their cellular response and the type of cell with which they interact. For example, the Bl chain peptide, YIGSR, does not interact with neuronal cells, whereas the A chain peptide, PA22-2, which contains SIKVAV, and the B2 chain peptide p20 recognize neuronal cells [ 1,3,4]. Likewise YIGSR, PDSGR and F9 (RYWLPR) are located on the Bl chain and inhibit tumor metastases [1,2,5,6,7-J], whereas PA22-2 increases tumor metastases, growth, angiogenesis, plasminogen activation and collagenase N activity [ 9,10,11**]. As YIGSR and SIKVAV have opposing activities in tumor metastases and angiogenesis, it is likely that the two sites are not available to cells in tissues at the same time, but rather are developmentally regulated by the presence of other matrix components. It is interesting to note that the all-D-con&tired IKVAV-containing peptide has similar biological activity to the al-L-IKVAV peptide [ 121. Various active peptides have been defined from the carboxyl globular domain of the A chain (termed G domain) and all appear to be active with neural cells [ 12,13*,14]. It should be noted that this domain has significant homology with other proteins including S protein, androgen-binding protein and sex hormone-binding globulin [ 15.1. An additional point to keep in mind is that some cells, such as endothelial cells, can recognize multiple sites on laminin including YIGSR, RGD and SIKVAV ( [ 16); HK Kleinman et al., unpublished data).
The extensive study of several adhesive macromolecules has been prompted by their widespread involvement in development and diseases. This review will describe the more abundant and better studied extracellular matrix components laminin, Iibronectin, collagens 1 and N, thrombospondin and entactin/nidogen, and their active sites defined to date by short synthetic peptides. Emphasis will be placed on the responding cells because active sites vary in cell types with different specificity. All of these components have been sequenced, which has facilitated the approach of employing synthetic peptides. Part of this review will focus on one of the laminin-derived synthetic peptides containing YIGSR (where YI is tyrile and GSR is from the single-letter amino acid code), which is more active in the polymeric form and has been found to block the differentiation of some cells, tumor growth and angiogenesis. Iaminin, tibronectin and the collagens contain multiple biologically active sites with distinct activities and cell specificities. In some cases, the cellular receptors for a particular site differ between specific cells, which leads to a large diversity of biological functions and cell regulation.
@
1992,
The MGSR site on the laminin Bl chain has been studied by several laboratories because of its potent inhibitory effect on tumor metastases [5,6,7*] and growth 1211. This peptide blocks tumor metastases by reducing tumor cell adhesion to laminin [4,22], but inhibits angiogenesis [7*,17,23]. The activity of this peptide has been increased by preparing it in the cyclic form, as a polymer, coupled to polyethylene glycol and as a ‘starburst’ structure [6,24,25]. It is not clear if the increased activity is due to a better conformation, a longer half-life Ltd
ISSN
0955-0674
819
820
Cell-to-cell
contact
and extracellular
matrix
. rable
1. Cell
attachment
tiolecule
sequences
in extracellular
matrix
Recognition (single-letter
-aminin
proteins. Cell
sequence amino
acid
type
distribution/function
code
Bl
YIGSR
Many
YICSR
cells,
RYVVLPR
F9
PDCSR
Synergistically
LCTIPC
Binds
PDGSR LCTIPC
site
inhibits
Heparin
metastasis
binding
functions
with
to eleastin
YICSR?
receptor
Laminin 82 RNIAEIIKDA
P20
Promotes
neurite
Ciliary
ganglion
outgrowth
Laminin s LRE site Laminin
LRE
neurons
A
PA22-2
IKVAV
Promotes
neurite
outgrowth,
activation RCD
site
Endothelial
RGDN
GD-1
KATPMLKMRTSFHCCIK
CD-2
KEGYKVRLDLNITLEFRUSK
CD-3
KNLEISRSTFDLLRNSYCVRK
CD-4
DGKWHTVKTEYIKRKAF
CD-6
KQNCLSSRASFRCCVRNLRLSR
and
Promote
plasminogen
tumor cells
neurite
formatlon and
others
outgrowth,
heparin
blnding i Melanoma
cells
Fibronectin RGD
site
RGDS
CSI
site
LDV
REDV
site
Synergestic
Most Most
cells,
neural
REDV site
Not
yet
inhibits
metastasis
crest-derived
cells,
Melanoma Most
determined
cells
Peptide
I
YEKPGSPPREVVPRPRPGV
Melanoma
cells
Peptide
II
KNNQKSEPLIGRKKT
Melanoma
cells
Collagen
IV
IV-H1 Hep
GVKGDKGNPGWPGAP Ill
Melanoma
GEFYFDLRLKGDK
cells,
promotes
Heparin
binding
Collagen
I
RGD
site
RGDTP
Fibroblasts
and
RGD
site
SRGRTG
Fibroblasts
and
DGEA
lymphocytes
cells
site
DGEA
Platelets
and
motility
others others others
Thorombospondin VTXG RGD
site site
WSXW Entactin RGD
VTXG
Promote
platelet
aggregation,
metastasis
RGD site
wsxw
Heparin
binding
(Nidogen) site
FRGDS
in the bloodstream, or a combination of both. Some difficulties have been encountered with the synthesis and stability of the pentameric molecule and this has led to some inconsistencies for its activity. The YIGSR peptide has also been found to interact with normal cells and to influence cell differentiation. As already mentioned, angiogenesis is blocked in zUt!o in the chick chorio-allantoic membrane and in a rabbit eye model; in addition, a specific stage of sea urchin embryonic development, archenteron elongation, is blocked while other stages are unaffected [ 70,261. The migration of endocardial cushion mesenchyme and chick neural crest from the notochord is also blocked by YIGSR
Mammary
tumor
cells
[27,28]. Many cells are observed to differentiate when plated on a Iaminin-rich basement membrane matrix [29]. In some, but not all, cases, YIGSR can block this differentiation. For example, Sertoli cell cord formation is blocked by MGSR due to a decreased cell attachment to the matrix [30]. Similar results are observed with bone cells [ 311, whereas endothelial cells remain attached to the matrix but do not align [ 161. In contrast, the formation of salivary gland-like structures on basement membrane is unaffected by MGSR, but is blocked by the SIKVAV-containing peptide [ 32.1. These &ta demonstrate that YIGSR is one of the major functional sites in laminin for a variety of cells.
Functional
Table 2. Activities
of laminin-derived
YIGSR peptide.
Activity
Cell type
Promotes
adhesion
HT1080,
or system
CHO,
endothelial
cells.
trophoblasts Blocks
laminin
adhesion
OVCAR-3,
colon
cell lines, PMN,
macrophages Promotes
migration
B16-FIO.
chick
endocardial
cushion
mesenchyme Inhibits
tumor
growth
lnhiblts
angiogenesis
B16-FlO,
Tsu-prl,
small
carcinoma, Chick
sarcoma
chorio-allantoic rabbit
cord
formation
Inhibits
urchin
Inhibits
outgrowth
development
vein
Primary Sea urchin
180 membrane,
eye model,
unbilical Inhibits
cell lung,
adhesion
molecules
Yamada and Kieinman
peptide blocks platelet adhesion to collagen, but not to fibronectin or laminin. Surprisingly, it also blocked T47D breast adenocarcinoma cell adhesion to collagen I and laminin. Both platelets and the T47D cells use the same receptor (azpI integrin) in their interaction with this sequence. Thus, the ability of cells to interact with this site is dependent on the presence of a specific receptor. The DGEA sequence recognizes the azpl integrin. This sequence is not in laminin, which suggests that a2pI can recognize additional sequences. Collagen N also has several active sites [2,42]. A major cell-binding site interacts with both the alpI integrin and the al& receptor [42,43**]. The specificity among various cell types has not yet been determined.
cells archenteron
elongation trunk
neural
crest
Fibronectin Fibronectin (molecular weight = 540000) is a serum protein that is also found in fibrous connective tissue and in the basement membrane of certain tissues [ 1,331. Like laminin, it has multiple active sites with different cellular specificities. The RGD sequence in libronectin was the first sequence to be reported active in promoting cell adhesion [34]. This sequence is found in more than 100 other proteins, including laminin, collagens I and N. entactin, vitronectin and fibrinogen [ 11. As some of these proteins are not biologically active, it would appear that the sequences and structure surrounding RGD are important for the activity in the native molecules. In fact, several upstream sequences in fibronectin have been reported to function synergistically with RGD in tibronectin [35-l. Additional active sites on fibronectin have been described, including LDV, which are cell-specific and in some cases located in alternatively spliced domains 136,371. All of the active sequences identified in libronectin have so far been found mainly to promote cell adhesion and sprcJding. The RGD site can reduce tumor metastases [ 38,391, but it is less active than the laminin-derived YIGSR.
Collagen
of cell
cell in vitro
Sertoli
embryo
Stage 12 chick
domains
I and IV
Collagens have been used to coat culture dishes because they promote cell adhesion and survival. Collagen I forms a Iibrillar network, often with fibronectin in L@ljo,whereas collagen N forms a complex non-Iibrillar network. Besides the RGD sites that are present on both collagens [2,-iO], type I collagen has a DGEA site, which is active with platelets and certain other cells [41**]. This
Thrombospondin
and entactin
(nidogen)
Thrombospondin is a large glycoprotein (molecular weight = 450 000) that is released from platelets. It is also present in various extracellular matrices due to its interactions with heparin, fibronectin and collagen. It has been found to promote cell adhesion and various other biological events [44,45-,46-l, including neurite outgrowth and inhibition of tumor metastases. To date, three cellbinding sites have been defined, and two of these are in the carboxyl-terminal heparin-binding domain. One peptide that is constructed from the type I repeat WSXW promotes melanoma cell adhesion, whereas another type I repeat peptide CSVTCG and its homologues both promote adhesion and inhibit platelet aggregation and tumor metastases [ 46.0,471. Entactin/nidogen (molecular weight = 158000) is found in all basement membranes, where it binds to other bioactive components including laminin and collagen N. It also binds well to collagen I. The RGD site on entactin is functional with various cells [43**,48-501. Due to its presence in complexes with other bioactive matrix molecules, it can be expected that some synergy in activity exists although that has not yet been demonstrated. The interaction of enactin with tibronogen chains also suggests a role in hemostasis [49].
Conclusions A number of cell adhesion molecules are multifunctional and demonstrate cell type specificity. Several active sites have been identified by synthetic peptides and others probably exist. The definition of active sites and duplication of their activities by small synthetic peptides has helped to unravel specific developmental mechanisms. Such peptides may also be used clinically in the treatment of diseases, clotting problems, or to coat prostheses and implants for better tissue adherence. These peptides may also be used to improve wound healing and/or reduce scarring.
821
822
Cell-to-cell
contact
References
and extracellular
and recommended
Papers of particular interest, review, have been highlighted . of special interest .. of outstanding interest 1.
matrix
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reading within
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period
of
chromoatogmphy,
1-l.
SK~~ISI~~ APN. LI’.TOIIRNEAI: PC, WAYNER E. FIII~CHT LT: Synthetic Peptides from the (&-boxy-terminal Clobulus Domain of the A Chain of Laminin Their Ability to Promote CeU Adhesion and Neurite Outgrowth and Interact with Heparin and the 81 lntegrin subunit. ./ CeN Biol 1991. 115:1137-l l-18.
1. ENGEL 1: Structure and Function of of a M&domain Protein. rASEl3.I 1990,
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3.
TASHIRO K, SEPHEI. GC, WEEKS BS. SASAKI M. MARTIN GR, KLEINMAN HK, YA~~ADA Y: A Synthetic Peptide Containing the IKVAV Sequence from the A Chain of Laminin Mediates Cell Attachment, Migration and Neurite Outgrowth. ./ Rid Chm 1989, 264:1617&16182.
4.
;finin tide.
Recognition
Sequences.
.I 13iol Chew
GRAF J. Iw~h!o~o Y. S*AKI M, MARTIN GR. KIEINMAN HK, Ro13n FA, YA.LMADA Y: Identification of an Amino Acid Sequence in Laminin Mediating Cell Attachment, Chemotaxis and Receptor Binding. GzN 1987, 48~989996.
integrin
ajpl
was found
to bind
to the pep-
15. .
Jos~:.I+I DR, BAKER ME: Sex Hormone Binding Globulin, Androgen Binding Protein and Vitamin K-dependent Protein S Are Homologous to Laminin A, Merosin. and Drosophila Crumbs Proteins. /+l.Sf?B ./ 1992. 6:21?7-2481. Strong homologies in the G domain (carboxTl terminus) of the A chain of Iaminin are obsencul nith various proteins. 16
GKANT
DS. T&+IIRO K. Secili-REAL B. YM~ADA I’, MAK~IN GR. I1K: Two Different Laminin Domains Mediate the Differentiation of Human Endothelial Cells into Capilliarylike Structures. Cc/I 1989, 58:933-943.
tiINhL.w
17.
SANES J, ENG\‘AU E, BLI~OWWI erogeneity of Basal Laminae: lagen IV at the Neuromuscular Cell Bid 1990, 111:1685-1699
18.
HL’NTER DD, SHAH V, MERUE JP. SANES JR: A Laminin-like Adhesive Protein Concentrated in the Synaptic Cleft of the Neuromuscular Junction. Nn!lrrr 1989, 338:223-23-i.
Effect of on the In1 ./ Rio/
19.
HIIN~L!R DD. POR-IZR BE, B~II~K JM. ADUIS SP. MER~E JP, SAIIES JR: Primary Sequence of a Motor Neuron Selective Adhesive Site in the Synaptic Basal Lamina Protein S-laminin. Cell 1989, 59905-913.
SAKA~~OTO N, IW’AH~A IM, TANS NG, OSADA Y: Inhibition of Angiogenesis and Tumor Growth by a Synthetic Laminin Peptide CDPGYIGSR-NH2. Cancer- Res 1331. 51:90~906. With an ID% of l.l3pg/egg, CDPGYIGSR blocks angiogenesis in the chick chorioallantoic membrane assay. Four days after intravenous tumor innoculation, daily injections of the peptide reduced tumor volume by more than 80%. The authors concluded that the peptide reduced tumor growth due to its anti-angiogenic eff‘ect.
20.
HUSKER DD, C~HXIAN N. MORRIS-V,UERO R, Bti~.oclc JW. AD,u.~s SP. SANES JR: An LRE (Leucine-Arginine-Glutamate)-dependent Mechanism for Adhesion of Neurons to S-laminin. J Newosci 1‘991, 12:396&3971.
5.
IWAMOTO
Y, MARTM of Iaminin 328:1132-l
6.
Y, Ro~t7’ FA, GR~ J. SUAKI M. KIJZIN~IAN HK, Y&\IAI)A GR: YIGSR a Pentapeptide from the Bl Chain Inhibits Tumor Cell Metastases. Science 1987, 134.
MURATA J, SAIKI 1. Azuhw I, NISHI N: Inhibitory a Synthetic Polypeptide, Poly (Tyr-Ue-Gly.Ser-Arg), Metastatic Formation of Malignant Tumor Cells. ilhxmo/ 1989, 11:92-99.
7.
.
8.
SKlrBrrz APN, ~McCAR~~~ JB, ZHOA Q. YI X. FIIRCHT LT: Definition of a Sequence RYVVLPR, within Laminin F-9 that Mediates Metastatic Fibrosarcoma CeU Adhesion and Spreading. Cancer Res 1990, 50~7612-7622.
9.
KANEMOTO T, REICH R. GREA~OP&X D. AOLER SH. YA~u~,A Y. KLEINMAN HK: Identification of an Amino Acid Sequence from the Laminin A Chain which Stimulates Metastases Formation and CoUagenase IV Production. Proc Nor/ Au& Sci USA 1990, 87:227’+2283.
10.
STACK S, GRAY RD, PKZO SV: Modulation of Plasminogen Activation and Type IV CoUagenase Activity by a Synthetic Peptide Derived from the Laminin A Chain. Biochetnimy 1990, 30:2073-2077.
11.
K~OTA S, TA~HIRO K, YA&IADA Y: Signaling Site of Laminin with Mitogenic Activity. J Bid chetn 1992, 267342854288. ze laminin A chain peptide containing SIKVAV stimulates PC12 cell groti and mRNA expression of c-/a\- and c-&n. The binding of both the-se proto-oncogene products to the API site m also increased b) the peptide. 12.
NOMIZU M, UTANI A, SHIRA~SHI N, KIBBEY MC, YAMADA Y, ROUER PP: The AU-D-COntiguratiOn Segment Containing the IKVAV Sequence of Laminin A Chain has Similar Activities to the AU-L-peptide in Vitro and in Vivo. J Biol C!wn 1932. 267:1411&14121.
21.
FR~D~IAN R. G~ACONNE G, K..mh!OTO T. MARTIN GR. G,~DAR AF, MLIISHINE JL: Reconstituted Basement Membrane (Matrigel) and Laminin Can Enhance the Tumorogenicity and Drug Resistance of Small CeU Lung Cancer Lines. Proc N&l Acad Sci I’SA 1990, 87:669ti707. Describes a new method for growing tumor cells that employs subcutaneous co-injection of tumor cells and basement membrdne. The peptide IlGSR blocks the growth of these tumors when co.injected whereas the peptide MGGR is without effect. 22.
DANEKEK DW, Pwzw AJ. SMELL: GD, MERC~IRIO AM: Relationship between Extracellular Matrix Interactions and Degree of Differentiation of Human Colon Carcinoma CeU Lines. Cuncer Res 1989, 496814.
23.
SWENR’
GEHEEN KR. SIURAMARAo P, FURCHT LT, SKLIBI-I-~ APN: A Synthetic Peptide Derived from the Carboxyl Terminus of the Laminin A Chain Represents a Biding Site for a$, Integrin. J Cell Biol 1992, 117:44’+459. A peptide from the globular domain of the laminin A chain supponed cell attachment and blocked laminin-mediated cell adhesion; antibody to the peptide also blocked laminin-mediated adhesion. Using peptide
TM, KIBBEI’ MC, ZAIN M, FwDhtU‘I R. KLEIN~WN HK: Membrane and the Laminin Peptide Containing Promote Tumor Growth and Metastases. Cancer Meluskzsis Reel 1991, 10:245-254. Basement SIKVAV
23.
KAW~AKI K. NA~~AKAWA M, MUMI T. M~;~AITA H, IWAI Y. Hahn\ T, MA~~I~II T: Amino Acids and Peptides XIV, Laminin Related Peptides and Their Inhibitory Effect on Metastases Formation. Bicdem Biopqa Res Commun 1991, 174:115%1162.
25.
KwNhlAh’ IHK, GRAI; J, IWAMOT’O Y. SASAKI M, SCHASTEEN CS, YMLADA Y. MARnN GR, RO~EY FA: Identification of a Sec. ond Active Site in Lam&n for Promotion of CeU Adhesion, Migration, and Inhibition of in Viuo Melanoma Lung Colonization. ArdJ Biocbem Bicgdga 1989. 272:39-45.
26.
BENX)N S, CHLI~PA S: DUTerentiation of in Vitro Sea Urchin Micromeres on Extracellular Matrix in the Absence of Serum. J Eq Zwl 1990, 256:222-226.
27.
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BILOZUR ME, HAY ED: Neural Crest Extracellular Matrix Utilizes Laminin, lagen. Deaf Biol 1988, 174:1159-1162.
13. .
K, HUNTER D: Molecular Hetlsoforms of Laminin and ColJunction and Elsewhere. J
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Functional 29.
KwNhwi-4 HK, GRAY J, IWAMOTO Y. K~I-~EN GT. om RC, SASAKI M, YAMAHA Y, MARTIN GR, LIICIXNI~IU-EDDS L: Role of Basement Membrane in Cell Differentiation. N I’ Acud Sci 1987. 513:134-145.
30.
HADLEY MA, WEEKS BS, KLEINMAN HK, motes Tubule Formation by Cultured 1990, 140:31%327.
31.
V~IKICRIC C, LLJITEN FP. KIEIN~WN HK. REDDI tiation of CanaUcuU-like Bone Cell Network Membrane Matrix Components and Defined Laminin. Gel/ 1990. 63:437-i45.
DYM M: Laminin ProSertoU Cells. Deft Riol
AH: Differenby Basement Domains of
32.
KIBH~Y MC, ROYCE LS, DYhi M, BALIM BJ. Ku1Nhi.m HK: Glandular Morphogenesis of a Human Submandibular Cell Line by Basement Membrane Components in V&o. E@ Ceil Kes 1992, 198:343-357. A human submandibular cell line forms glandularlike structures on a basement membmne substrate. The laminin peptides MGSR and RGD have no effect on the morphology of the cells, whereas the SIKVAV pep tide blocks gland formation and causes a rapidly growing monolayer of cells to form. .
33.
PIERXHBACHER of Fibronectin merits of the
34.
YUIADA proteins. by Hay
KM: In E.D.
MD, RIIOSWHTI E: Cell Attachment Activity Can be Duplicated by Small Synthetic Frag Molecule. Nntfrre 1984. 309:30-33. Fibronectin Cell Bidon? New York:
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Other Cell Interactive of fl,e I:v!rucell~rlcrr .Ilrr/ris Plenum Press; 1991:111-l-16.
GlycoEdited
AOTA S, NACAI T, YA~MDA KM: Characterization of Regions of Fibronectin besides the Arginine-Glycine-Aspartic Acid Sequence Required for Adhesive Function of the Cell-binding Domain Using Site-directed Mutagenesis. ./ Bid Cheer 1991, 266:1593%15943. Using deletion analysis and oligonucleotide site-directed mutagenesis, regions on libronectin were found to be required for full activity of the RGD site.
35.
..
36.
37.
HLlhIPHlUES MJ, kCIYAh!A SK, KOhlORlYA A, OlDEN K, \I’A!\lAI,A KM: Identification of an Alternatively Spliced Site in Human Plasma Fibronectin that Mediates CeU Type-specific Adhesion. J Cell Rio/ 1986. 103:2637-2(~7. HLIhwHms MJ. KO~IOR~‘A A, AK~‘~IA SK, OIIXIN K. Y.~\!AI)A KM: Identification of Two Distinct Regions of the Type 111 Connecting Segment of Human Plasma Fibronectin that Promote CeU Type-specific Adhesion. J Hi01 C/~em 1989,
domains
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40.
of the Peptides Murine
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adhesion
Yamada
molecules
ST’AATZ WD. Foli KF, ZULTER MM, SANr’oTo SA: Identification of a Sequence for the a,Pl Integrin 1991, 2667363-7367. Peptides preparcil from cyanogen bromide lagen identilird a four amino acid sequence and other cells in their a#, integrin-mediated also blocked T47D breast adenocarcinoma
?l. ..
42.
and Kleinman
ADA\IS SP, RODRIQIIU. BA, Tetrapeptide Recognition in Collagen. J No/ Cbem fmgment CB3 of type 1 col( DGEA) used by platelets adhesion, This peptide adhesion to laminin.
CHEIXRC MK, hlcC,%aT!n’ JB, SI(t!BrrL APN. FL~RCHT LT. TSILIHARS EC: Characterization of a Type IV Collagen that Promotes Melanoma CeU Adhesion, Spreading and Mobility. ./ CeN Rio1 1991. 111:201-270.
t3.
VANLIENBERC P. KEKN A, RIES A, LUCKENBIU.EDDS I+ WN K, K~IHN K: Characterization of a Type N Collagen Major CeU Binding Site with Affinity to the A, B, and the &PI lntegrins. J Cell Biol 1991, 113:1475-1483. A cyanogen bromide fragment CB3 of collagen IV is ils active as the intact molecule in promoting cell adhesion. Using CB3 affinity chromatogmphy. two integrins. alal and a&- - were isolated. ..
i-1.
Trlsn?ls~~ L. Shim-1 Promoted 236: 15X-
GP. ROlJlhtiN Vl., Mt’lU’~ A, SlEGlIR K. ShllTH S. KORIXWSKI J, L~LIDSEN KA: Thrombospondin CeU Substratum Adhesion. Science 1987, 1573.
45. ..
O’SHM KS, LLII L-H. DIN-~ VM: Thrombospondin and a 140 kD Fragment Promote Adhesion and Neurite Outgrowth from Embryonic Central and Peripheral Neurons and from PC12 Cells. Neurojr 1991, 7:231-237. Neurite outgrowth did not involve the RGD sequence or the aminoterminal heparin.binding domain.
-16.
TI’S~YNSKI GP. Ro7HhIAN VL, DELIXH AH, HA~IILI’ON BK, EYAL J: Biological Activities of Peptides and Peptide Analogues Derived from Common Sequences Present in Thrombospondin, Properdin, and Malarial Proteins. J Cell Bioll992, 1 16:209-2 17. Thrombospondin is produced by platelets and influences platelet aggre+on, cell adhesion and tumor metastases. Two active sites, which are also present in a number of other proteins including properdin and malarial proteins. contain the acthig. ..
-i7.
GL,O N. KHI’T~XH, NEC,II~: E, VOGEI. T. 1~~ D& ROBERIS DD: Heparin and Sulfatide-binding Peptides from the Type I Repeats of Human Thrombospondin Promote Melanoma CeU Adhesion. Proc N&l Auzrl Sci I’SA 1992. 89:304&3013.
ix.
OIAKR%\‘AHTI S. T~L\I MF. CHEIINC; AE: The Basement Membrane Glycoprotein Entactin Promotes CeU Attachment and Binds Calcium Ions. J Biol C/Jet,? 1990, 265:1059?-10603.
-19.
,~L!NN K, DEIVJIANN R. A~:DWI.IEY M. TNPL R, RNhIoml L, YMIADA Y. P&v TC. CohwAY D. CHI: ML: Amino Acid Sequence of Mouse Nidogen, a Multidomain Basement Membrane Protein with Binding Activity for Laminin. Collagen N and Cells. EVBO J 1989. 8:65-‘7.
50.
Wr’ CY. CHL:NG AE: Potential Role of Entactin in Hemostasisspecific Interactions of Entactin with Fibrinogen A-a and B-P Chains. ,/ Rio/ (3enr 1991, 266:18802-18807.
262:68866892. 38.
of cell
Y Yamdda and HK KleinnXIn. laboratory National Institute of Dental Rt!se’drch. Bethesda, Mar&and 20892, USA
of Developmental National Institutes
of
Biology, Health.
823