Enamelysin

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

[26] van Horssen, J., Vos, C.M., Admiraal, L., van Haastert, E.S., Montagne, L., van der Valk, P., de Vries, H.E. (2006). Matrix metalloproteinase-19 is highly expressed in active multiple sclerosis lesions. Neuropathol. Appl. Neurobiol. 32, 585
593. [27] Sadowski, T., Dietrich, S., Muller, M., Havlickova, B., Schunck, M., Proksch, E., Muller, M.S., Sedlacek, R. (2003). Matrix metalloproteinase-19 expression in normal and diseased skin: dysregulation by epidermal proliferation. J. Invest. Dermatol. 121, 989
996. [28] Suomela, S., Kariniemi, A.L., Impola, U., Karvonen, S.L., Snellman, E., Uurasmaa, T., Peltonen, J., Saarialho-Kere, U. (2003). Matrix metalloproteinase-19 is expressed by keratinocytes in psoriasis. Acta Derm. Venereol. 83, 108
114. [29] Jain, S., Suarez, A.A., McGuire, J., Liapis, H. (2007). Expression profiles of congenital renal dysplasia reveal new insights into renal development and disease. Pediatr. Nephrol. 22, 962
974. [30] Johnston, P., Larson, D., Clark, I.M., Chojnowski, A.J. (2008). Metalloproteinase gene expression correlates with clinical outcome in Dupuytren’s disease. J. Hand Surg. Am. 33, 1160
1167. [31] Kuivanen, T., Tanskanen, M., Jahkola, T., Impola, U., AskoSeljavaara, S., Saarialho-Kere, U. (2004). Matrilysin-1 (MMP-7) and MMP-19 are expressed by Paget’s cells in extramammary Paget’s disease. J. Cutan. Pathol. 31, 483
491. [32] Baum, O., Ganster, M., Baumgartner, I., Nieselt, K., Djonov, V. (2007). Basement membrane remodeling in skeletal muscles of patients with limb ischemia involves regulation of matrix

[33]

[34]

[35]

[36]

[37] [38]

[39]

835

metalloproteinases and tissue inhibitor of matrix metalloproteinases. J. Vasc. Res. 44, 202
213. Shin, J.I., Song, K.S., Kim, H., Cho, N.H., Kim, J., Kim, H.S., Lee, J.S. (2011). The gene expression profile of matrix metalloproteinases and their inhibitors in children with Henoch-Schonlein purpura. Br. J. Dermatol. 164, 1348
1355. Lohi, J., Wilson, C.L., Roby, J.D., Parks, W.C. (2001). Epilysin, a novel human matrix metalloproteinase (MMP-28) expressed in testis and keratinocytes and in response to injury. J. Biol. Chem. 276, 10134
10144. Werner, S.R., Mescher, A.L., Neff, A.W., King, M.W., Chaturvedi, S., Duffin, K.L., Harty, M.W., Smith, R.C. (2007). Neural MMP-28 expression precedes myelination during development and peripheral nerve repair. Dev. Dyn. 236, 2852
2864. Davidson, R.K., Waters, J.G., Kevorkian, L., Darrah, C., Cooper, A., Donell, S.T., Clark, I.M. (2006). Expression profiling of metalloproteinases and their inhibitors in synovium and cartilage. Arthritis Res. Ther. 8, R124. Werner, S.R., Dotzlaf, J.E., Smith, R.C. (2008). MMP-28 as a regulator of myelination. BMC Neurosci. 9, 83. Rodgers, U.R., Kevorkian, L., Surridge, A.K., Waters, J.G., Swingler, T.E., Culley, K., Illman, S., Lohi, J., Parker, A.E., Clark, I.M. (2009). Expression and function of matrix metalloproteinase (MMP)-28. Matrix Biol. 28, 263
272. Illman, S.A., Lehti, K., Keski-Oja, J., Lohi, J. (2006). Epilysin (MMP-28) induces TGF-beta mediated epithelial to mesenchymal transition in lung carcinoma cells. J. Cell Sci. 119, 3856
6385.

Miriam Fanjul-Ferna´ndez Departamento de Bioquı´mica y Biologı´a Molecular, Instituto Universitario de Oncologı´a, IUOPA, Universidad de Oviedo, 33006-Oviedo, Spain.

Carlos Lo´pez-Otı´n Departamento de Bioquı´mica y Biologı´a Molecular, Instituto Universitario de Oncologı´a, IUOPA, Universidad de Oviedo, 33006-Oviedo, Spain. Handbook of Proteolytic Enzymes, 3rd Edn ISBN: 978-0-12-382219-2

© 2013 Elsevier Ltd. All rights reserved. DOI: http://dx.doi.org/10.1016/B978-0-12-382219-2.00170-8

Chapter 171

Matrix Metalloproteinase-20/Enamelysin DATABANKS

Name and History

MEROPS name: matrix metallopeptidase-20 MEROPS classification: clan MA, subclan MA(M), family M10, subfamily M10A, peptidase M10.019 Species distribution: superclass Tetrapoda Reference sequence from: Homo sapiens (UniProt: O60882)

Matrix metalloproteinase-20 (enamelysin, MMP20) is so named because it was originally isolated by PCR-based homology cloning from a porcine enamel organ-specific cDNA library. At the time of cloning, the deduced amino acid sequence had the highest identity (49%) to porcine interstitial collagenase (matrix metalloproteinase-1;

836

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

Chapter 152). Enamelysin was not detected in several other tissues assayed by northern blot analysis and since the enamel organ is responsible for dental enamel formation, enamelysin seemed an appropriate name for a toothspecific matrix metalloproteinase (MMP) [1]. Prior to the identification of MMP20, an unidentified metalloproteinase(s) was observed by performing inhibitor/zymography studies on extracted enamel proteins [2,3]. Two of these metalloproteinase bands were later purified from the enamel, sequenced and identified as the N-terminus of the protein encoded by the previously cloned Mmp20 transcript [4]. After the original identification of MMP20, the ameloblasts of the enamel organ and the odontoblasts of the dental papilla were demonstrated as positive for MMP20 expression [5
7]; however, MMP20 expression proved to be highly restricted. In one study, 51 different cell lines were assessed by RT-PCR for MMP20 expression, but none of the cell lines tested positive [8]. Even so, MMP20 expression was observed in pathologic tissues such as in ghost cells of calcifying odontogenic cysts [9], odontogenic tumors [10], human tongue carcinoma cells [11] and in bradykinin treated granulosa cells isolated from the follicles of porcine ovaries [12]. A comprehensive assessment of MMP20 expression by qPCR of mRNA isolated from various mouse tissues revealed that other than the developing tooth, MMP20 was only expressed at very low levels in the large intestine [13]. The expression levels in the intestine were so low they were undetectable by Northern blot analysis and were approximately 5000 times lower than the levels observed in 4-day-old tooth buds. MMP20 was not expressed in small intestine, brain, heart, kidney, liver, lung, pancreas, spleen, or stomach [13]. Baleen whales are filter feeders that lack teeth and a recent report showed that Mmp20 is a pseudogene in these whales. Because nonfunctional Mmp20 genes are found only in mammals lacking enamel, the authors suggested that the only non-overlapping function of MMP20 is in dental enamel formation [14]. Therefore to date, MMP20 is considered a tooth-specific MMP.

Activity and Specificity Mmp20 was originally cloned from the enamel organ [1] and was first localized to the forming enamel [4,7]. Several studies demonstrated that MMP20 cleaves the most abundant enamel matrix protein, amelogenin [5,7,15
19]. Two different studies identified the exact MMP20 cleavage sites in amelogenin. The first study used recombinant proteins [18] and the second used native amelogenin and native MMP20 purified from developing pig teeth [19]. The precise cleavage sites were identified by various means including mass spectrometry

and protein sequencing. These cleavage sites were then compared to previously identified amelogenin cleavage products isolated from extracted porcine enamel. All of the MMP20 amelogenin cleavage sites generated in vitro were also identified from amelogenins extracted from normal porcine enamel. Subsequently, the other enamel matrix proteins, ameloblastin, enamelin, and kallikrein-4 (KLK4) were also assessed to determine if they are MMP20 substrates. MMP20 was demonstrated to cleave the KLK4 propeptide to produce catalytically active KLK4 [20]. Similar exacting protocols used to identify amelogenin cleavage products were also used to identify ameloblastin and enamelin cleavage products. Like amelogenin, all of the ameloblastin MMP20 cleavage sites identified in vitro were also identified from ameloblastin extracted from porcine enamel in vivo [21,22]. Enamelin has a 186 kDa apparent molecular weight and is highly glycosylated. However, only a 32 kDa enamelin cleavage product accumulates within the maturing subsurface enamel layer [23]. MMP20 cleaved the 32 kDa enamelin only after it was deglycosylated [24]. Therefore, MMP20 is likely responsible for generating the 32 kDa enamelin cleavage product in vivo. Also, MMP20 and MMP2 were each demonstrated to cleave dentine sialophosphoprotein, which is the major noncollagen secretory product of odontoblasts responsible for dentin formation [25]. Taken together, these results highlight MMP20 as the predominant enamel matrix processing enzyme during the secretory stage of enamel formation. In addition to enamel and dentin proteins, MMP20 was also demonstrated to cleave casein and/or gelatin [4,7,11], aggrecan and cartilage oligomeric matrix protein [26], type V collagen [13], type XVIII collagen [27], fibronectin, type IV collagen, tenascin-C, laminin-1 and -5, but not type I or type II collagen [11]. Recombinant MMP20 autoactivates [16] and appears to readily remove its hemopexin-like domain to form a catalytically active species of approximately 22
27 kDa [4,15,18]. Two different studies have determined kcat/Km values for recombinant human or porcine MMP20 based on results from incubation with quenched fluorescent peptides. Despite the use of MMP20 from different species, and the use of different peptides, the values were in good agreement ranging from 4.40
5.88 3 104 M21 s21 [16,18]. One study characterized MMP20 substrate specificity by use of an iterative mixture-based random doedecamer peptide library screen with Edman sequencing of MMP20 cleavage products. It was determined that MMP20 has a broad substrate specificity with a deep and wide catalytic pocket that can accommodate substrates with large aromatic residues in the P10 position.

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

This study suggested that MMP20 expression may be restricted to tooth tissues because of its broad substrate specificity that might otherwise cause tissue destruction if expressed elsewhere [13].

Structural Chemistry MMP20 shares a domain structure characteristic of the MMPs. MMP20 has a signal peptide, a propeptide, a catalytic domain, and a hemopexin-like domain. The MMP20 preproprotein is 483 amino acids in length and has a predicted Mr of 54 100. The proprotein is 461 amino acids in length with a predicted Mr of 51 900 and removal of the 85 residue propeptide generates an active proteinase (42.5 kDa) composed of 376 amino acids. A tyrosine residue is at the N-terminus of the active enzyme in pig [1], human [16], cow [4], and mouse [6]. The NMR structure of the MMP20 catalytic domain complexed with N-isobutyl-N-(4-methoxyphenylsulfonyl) glycyl hydroxamic acid (NNGH) was reported and the NNGH adduct structure was most similar to MMP3-, MMP10- and MMP12-NNGH [28]. However, despite a domain structure similar to that of other MMPs, MMP20 also possesses several unique structural features that define it as a novel MMP. First, the MMP20 amino acid sequence contains no consensus N-linked glycosylation sites (Asn-Tyr-Ser/Thr). Other MMPs of a similar Mr to MMP20 (collagenases, metalloelastase, stromelysins-1 and 2) have conserved N-linked glycosylation sites within their catalytic domains [5,16]. Second, MMP20 lacks two of the three residues important in defining the active site of a collagenase and lacks all three of the residues important in defining the active site of a stromelysin [1,6,16]. Third, MMP20 has a unique hinge region that connects the catalytic domain to the hemopexin-like domain. The collagenases have 16residue hinge regions [29], the stromelysins have a 25-residue hydrophobic hinge region, and MMP-19 has a 32-residue acidic hinge region [30]. In contrast, MMP20 contains a 24-residue hinge region with a basic charge [1,4
6,16]. Thus, although MMP20 has a similar domain structure and Mr to the collagenases, stromelysins, and metalloelastase, it is not a member of any one of these groups.

Preparation Human [16], porcine [31], and bovine [15] MMP20 were each expressed in E. coli and purified by means of an introduced N-terminal His-tag. The full-length refolded enzyme is active, but tends to undergo autocatalysis to an active 22 kDa species.

837

Preparation of MMP20 from Mouse Tooth Germs The first molar was extracted from a 3
5-day-old mouse pup. The pulp was removed, and the crown was demineralized with 0.17 N HCl/0.98% formic acid containing protease inhibitor cocktail set III (Calbiochem) and 1 mM 1,10-phenanthroline for 2 h at 4 C (2 ml of demineralization solution incubated with 50 mg teeth). This was centrifuged for 10 min at 5000 rpm (6076 g) at 4 C. The supernatant was neutralized by dialyzing against 50 mM Tris-HCl buffer (pH 7.4) containing protease inhibitor cocktail set III. To accomplish this, the supernatant was removed (the pellet discarded) and the supernatant placed in a Spectra/Por dialysis tube/bag (MW cutoff 3500) and dialyzed against 1 liter of Tris-HCL buffer for 3 h at room temperature (dialysis was performed in a flask or beaker with stirring). The dialyzed supernatant was centrifuged for 10 min at 12 000 rpm (19 837 g) at room temperature; MMP20 was present in the pellet. Samples were dissolved with sample buffer for SDS-PAGE (100 µg/ 50 µl). For immunoblots and zymograms (Do Not Boil), 5
10 µl were run per lane (personal communication, Dr. Yasuo Yamakoshi, University of Michigan).

Preparation of MMP20 from Pig Tooth Germs Tooth germs of permanent molars are surgically extracted from the maxillae and mandibles of 6-month-old pigs. The enamel organ epithelia and dental pulp tissue are removed by means of tissue forceps. The soft, cheese-like enamel is separated from the crown with a spatula. Enamel scrapings are sequentially extracted to obtain neutral and alkaline extracts. The neutral extract (supernatant) is obtained by homogenization of the enamel samples in So¨rensen buffer (made by mixing Na2HPO4 and KH2PO4 to achieve a final phosphate concentration of 50 mM and a pH of 7.4), followed by centrifugation. The pellet is re-suspended in carbonate buffer (made by mixing NaHCO3 and Na2CO3 to a final concentration of 50 mM and a pH of 10.8), homogenized, and then centrifuged. This supernatant is designated as alkaline extract. The soft enamel-alkaline extract is fractionated by ion exchange chromatography in a Q-Sepharoset Fast Flow column (1.6 cm 3 20 cm, GE Healthcare Biosciences, Little Chalfont, UK) equilibrated with 50 mM Tris-HCl/ 6 M urea (pH 7.4) and eluted with a linear NaCl gradient (0.0
0.5 M). MMP20 elutes in the fourth peak and is desalted and concentrated by ultra-filtration (Ultracel; Millipore Corporation) and lyophilized. The lyophilized sample is dissolved with 20 mM sodium citrate/6 M urea (pH 7.4) and applied to a Heparin Sepharoset 6 Fast

838

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

Flow column (1.6 cm 3 20 cm, GE Health-care) equilibrated with the same buffer and eluted with a step NaCl gradient (0.02, 0.05, and 0.5 M at a flow rate of 0.2 mL/ min). MMP20 elutes in the second peak (i.e. 0.02 M NaCl), which is then buffer-exchanged with 50 mM TrisHCl (pH 7.4) with the use of a regenerated cellulose membrane (Ultracel; NMWL: 3000; Millipore) [19]. Active recombinant human matrix metalloproteinase20 (MMP20, enamelysin) produced in E. coli is commercially available (ENZO Life Sciences, Plymouth Meeting, PA). The enzyme consists of residues Tyr108-Phe276 (NM 004771), which comprises the catalytic domain of human MMP20. This is the active form of MMP20, which lacks the C-terminal hemopexin domain, but also has a C-terminal purification tag.

Biological Aspects The human MMP20 gene maps to the MMP cluster on chromosome 11q22.3 [16] and the mouse gene maps within the MMP gene cluster at the centromeric end of chromosome 9 [32]. During tooth development MMP20 transcripts are expressed by preameloblasts as the predentin is beginning to form and expression continues throughout the secretory, transition, and early maturation stages of enamel development [33]. However, during mid-maturation and into the late maturation stages, MMP20 expression is almost completely eliminated [5]. A membrane type-MMP (MT1-MMP, MMP-14) was identified on the cell surface of ameloblasts and odontoblasts of the developing tooth [34] and MT1-MMP was demonstrated to activate the MMP20 zymogen [35]. Homozygous deletion of the Mmp20 catalytic domain from the mouse genome resulted in a phenotype limited to dental enamel. The Mmp20 null mouse did not process amelogenin properly, possesses an altered enamel protein and associated enamel rod pattern, had hypoplastic enamel, had enamel that fractured from the dentin, and had a deteriorating tooth morphology as enamel development progressed [36]. These results suggested that one form of the enamel-specific human disease amelogenesis imperfecta (AI) may be caused by the recessively inherited inactivation of the MMP20 locus. Subsequently, four different MMP20 mutations were reported that each cause a severe form of autosomal recessive AI [37
40]. Zymography and Western blots demonstrate two different MMP20 bands of approximately 46 and 41 kDa [7]. The identity of the 46 and 41 kDa forms of purified porcine MMP20 was assessed by performing immunoblots, zymography, reverse-phase HPLC and protein sequencing after exposure to oxidizing and reducing conditions [41]. Under oxidizing conditions the disulfide bridge connecting the first and last amino acids of the

C-terminal hemopexin domain remains intact, and both the 46 and 41 kDa forms of MMP-20 were observed. Under reducing conditions, the disulfide bond is released and the 41 kDa band is replaced by a catalytically active 27 kDa band. Edman degradation of the three bands showed they all contained the catalytic domain at their Ntermini (YRLFPGEPK), proving that none of the bands corresponded to the MMP20 zymogen. In addition, under reducing conditions, a 17 kDa protein band stained positive for MMP20 on the immunoblot and its N-terminal sequence started with Ile336 of the hemopexin domain. Taken together, these observations demonstrate that one of the 46 and 41 kDa MMP20 bands is the active intact protease, while the other band is active protease that has been cleaved in the hemopexin domain after Thr335 [41]. This C-terminal peptide is covalently attached by the disulfide bridge. The MMP20 zymogen was not observed, presumably due to its efficient activation in vivo. [41].

Further Reading For reviews on the role of proteases in developing dental enamel refer to the following articles [42
44].

References [1] Bartlett, J.D., Simmer, J.P., Xue, J., Margolis, H.C., Moreno, E.C. (1996). Molecular cloning and mRNA tissue distribution of a novel matrix metalloproteinase isolated from porcine enamel organ. Gene 183(1
2), 123
128. [2] DenBesten, P.K., Heffernan, L.M. (1989). Separation by polyacrylamide gel electrophoresis of multiple proteases in rat and bovine enamel. Arch. Oral Biol. 34(6), 399
404. [3] Overall, C.M., Limeback, H. (1988). Identification and characterization of enamel proteinases isolated from developing enamel. Amelogeninolytic serine proteinases are associated with enamel maturation in pig. Biochem. J. 256(3), 965
972. [4] Den Besten, P.K., Punzi, J.S., Li, W. (1998). Purification and sequencing of a 21 kDa and 25 kDa bovine enamel metalloproteinase. Eur. J. Oral Sci. 106(Suppl. 1), 345
349. [5] Bartlett, J.D., Ryu, O.H., Xue, J., Simmer, J.P., Margolis, H.C. (1998). Enamelysin mRNA displays a developmentally defined pattern of expression and encodes a protein which degrades amelogenin. Connect. Tissue Res. 39(1
3), 101
109. [6] Caterina, J., Shi, J., Sun, X., Qian, Q., Yamada, S., Liu, Y., Krakora, S., Bartlett, J.D., Yamada, Y., Engler, J.A., BirkedalHansen, H., Simmer, J.P. (2000). Cloning, characterization, and expression analysis of mouse enamelysin. J. Dent. Res. 79(9), 1697
1703. [7] Fukae, M., Tanabe, T., Uchida, T., Lee, S.K., Ryu, O.H., Murakami, C., Wakida, K., Simmer, J.P., Yamada, Y., Bartlett, J.D. (1998). Enamelysin (matrix metalloproteinase-20): localization in the developing tooth and effects of pH and calcium on amelogenin hydrolysis. J. Dent. Res. 77(8), 1580
1588. [8] Grant, G.M., Giambernardi, T.A., Grant, A.M., Klebe, R.J. (1999). Overview of expression of matrix metalloproteinases (MMP-17,

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

[9]

[10]

[11]

[12]

[13]

[14]

[15]

[16]

[17]

[18]

[19]

[20]

[21]

[22]

MMP-18, and MMP-20) in cultured human cells. Matrix Biol. 18 (2), 145
148. Takata, T., Zhao, M., Nikai, H., Uchida, T., Wang, T. (2000). Ghost cells in calcifying odontogenic cyst express enamel-related proteins. Histochem. J. 32(4), 223
229. Takata, T., Zhao, M., Uchida, T., Wang, T., Aoki, T., Bartlett, J.D., Nikai, H. (2000). Immunohistochemical detection and distribution of enamelysin (MMP-20) in human odontogenic tumors. J. Dent. Res. 79(8), 1608
1613. Vaananen, A., Srinivas, R., Parikka, M., Palosaari, H., Bartlett, J.D., Iwata, K., Grenman, R., Stenman, U.H., Sorsa, T., Salo, T. (2001). Expression and regulation of MMP-20 in human tongue carcinoma cells. J. Dent. Res. 80(10), 1884
1889. Kimura, A., Kihara, T., Ohkura, R., Ogiwara, K., Takahashi, T. (2001). Localization of bradykinin B(2) receptor in the follicles of porcine ovary and increased expression of matrix metalloproteinase-3 and -20 in cultured granulosa cells by bradykinin treatment. Biol. Reprod. 65(5), 1462
1470. Turk, B.E., Lee, D.H., Yamakoshi, Y., Klingenhoff, A., Reichenberger, E., Wright, J.T., Simmer, J.P., Komisarof, J.A., Cantley, L.C., Bartlett, J.D. (2006). MMP-20 is predominately a tooth-specific enzyme with a deep catalytic pocket that hydrolyzes type V collagen. Biochemistry 45(12), 3863
3874. Meredith, R.W., Gatesy, J., Cheng, J., Springer, M.S. (2011). Pseudogenization of the tooth gene enamelysin (MMP20) in the common ancestor of extant baleen whales. Proc. Biol. Sci. 278 (1708), 993
1002. Li, W., Machule, D., Gao, C., DenBesten, P.K. (1999). Activation of recombinant bovine matrix metalloproteinase-20 and its hydrolysis of two amelogenin oligopeptides. Eur. J. Oral Sci. 107(5), 352
359. Llano, E., Pendas, A.M., Knauper, V., Sorsa, T., Salo, T., Salido, E., Murphy, G., Simmer, J.P., Bartlett, J.D., Lopez-Otin, C. (1997). Identification and structural and functional characterization of human enamelysin (MMP-20). Biochemistry 36(49), 15101
15108. Moradian-Oldak, J., Jimenez, I., Maltby, D., Fincham, A.G. (2001). Controlled proteolysis of amelogenins reveals exposure of both carboxy- and amino-terminal regions. Biopolymers 58(7), 606
616. Ryu, O.H., Fincham, A.G., Hu, C.C., Zhang, C., Qian, Q., Bartlett, J.D., Simmer, J.P. (1999). Characterization of recombinant pig enamelysin activity and cleavage of recombinant pig and mouse amelogenins. J. Dent. Res. 78(3), 743
750. Nagano, T., Kakegawa, A., Yamakoshi, Y., Tsuchiya, S., Hu, J.C., Gomi, K., Arai, T., Bartlett, J.D., Simmer, J.P. (2009). Mmp-20 and Klk4 cleavage site preferences for amelogenin sequences. J. Dent. Res. 88(9), 823
828. Ryu, O., Hu, J.C., Yamakoshi, Y., Villemain, J.L., Cao, X., Zhang, C., Bartlett, J.D., Simmer, J.P. (2002). Porcine kallikrein-4 activation, glycosylation, activity, and expression in prokaryotic and eukaryotic hosts. Eur. J. Oral Sci. 110(5), 358
365. Iwata, T., Yamakoshi, Y., Hu, J.C., Ishikawa, I., Bartlett, J.D., Krebsbach, P.H., Simmer, J.P. (2007). Processing of ameloblastin by MMP-20. J. Dent. Res. 86(2), 153
157. Chun, Y.H., Yamakoshi, Y., Yamakoshi, F., Fukae, M., Hu, J.C., Bartlett, J.D., Simmer, J.P. (2010). Cleavage site specificity of MMP-20 for secretory-stage ameloblastin. J. Dent. Res. 89(8), 785
790.

839

[23] Hu, J.C., Yamakoshi, Y. (2003). Enamelin and autosomal-dominant amelogenesis imperfecta. Crit. Rev. Oral Biol. Med. 14(6), 387
398. [24] Yamakoshi, Y., Hu, J.C., Fukae, M., Yamakoshi, F., Simmer, J.P. (2006). How do enamelysin and kallikrein 4 process the 32-kDa enamelin? Eur. J. Oral Sci. 114(Suppl. 1), 45
51. [25] Yamakoshi, Y., Hu, J.C., Iwata, T., Kobayashi, K., Fukae, M., Simmer, J.P. (2006). Dentin sialophosphoprotein is processed by MMP-2 and MMP-20 in vitro and in vivo. J. Biol. Chem. 281(50), 38235
38243. [26] Stracke, J.O., Fosang, A.J., Last, K., Mercuri, F.A., Pendas, A.M., Llano, E., Perris, R., Di Cesare, P.E., Murphy, G., Knauper, V. (2000). Matrix metalloproteinases 19 and 20 cleave aggrecan and cartilage oligomeric matrix protein (COMP). FEBS Lett. 478 (1
2), 52
56. [27] Vaananen, A., Tjaderhane, L., Eklund, L., Heljasvaara, R., Pihlajaniemi, T., Herva, R., Ding, Y., Bartlett, J.D., Salo, T. (2004). Expression of collagen XVIII and MMP-20 in developing teeth and odontogenic tumors. Matrix Biol. 23(3), 153
161. [28] Arendt, Y., Banci, L., Bertini, I., Cantini, F., Cozzi, R., Del Conte, R., Gonnelli, L. (2007). Catalytic domain of MMP20 (Enamelysin)
the NMR structure of a new matrix metalloproteinase. FEBS Lett. 581(24), 4723
4726. [29] Freije, J.M., Diez-Itza, I., Balbin, M., Sanchez, L.M., Blasco, R., Tolivia, J., Lopez-Otin, C. (1994). Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas. J. Biol. Chem. 269(24), 16766
16773. [30] Pendas, A.M., Knauper, V., Puente, X.S., Llano, E., Mattei, M.G., Apte, S., Murphy, G., Lopez-Otin, C. (1997). Identification and characterization of a novel human matrix metalloproteinase with unique structural characteristics, chromosomal location, and tissue distribution. J. Biol. Chem. 272(7), 4281
4286. [31] Fukae, M., Tanabe, T. (1998). Degradation of enamel matrix proteins in porcine secretory enamel. Connect. Tissue Res. 39(1
3), 123
129. [32] Caterina, J., Shi, J., Krakora, S., Bartlett, J.D., Engler, J.A., Kozak, C.A., Birkedal-Hansen, H. (1999). Isolation, characterization, and chromosomal location of the mouse enamelysin gene. Genomics 62(2), 308
311. [33] Begue-Kirn, C., Krebsbach, P.H., Bartlett, J.D., Butler, W.T. (1998). Dentin sialoprotein, dentin phosphoprotein, enamelysin and ameloblastin: tooth-specific molecules that are distinctively expressed during murine dental differentiation. Eur. J. Oral Sci. 106(5), 963
970. [34] Caron, C., Xue, J., Bartlett, J.D. (1998). Expression and localization of membrane type 1 matrix metalloproteinase in tooth tissues. Matrix Biol. 17(7), 501
511. [35] Palosaari, H., Ding, Y., Larmas, M., Sorsa, T., Bartlett, J.D., Salo, T., Tjaderhane, L. (2002). Regulation and interactions of MT1-MMP and MMP-20 in human odontoblasts and pulp tissue in vitro. J. Dent Res. 81(5), 354
359. [36] Caterina, J.J., Skobe, Z., Shi, J., Ding, Y., Simmer, J.P., BirkedalHansen, H., Bartlett, J.D. (2002). Enamelysin (matrix metalloproteinase 20)-deficient mice display an amelogenesis imperfecta phenotype. J. Biol. Chem. 277(51), 49598
49604. [37] Kim, J.W., Simmer, J.P., Hart, T.C., Hart, P.S., Ramaswami, M.D., Bartlett, J.D., Hu, J.C. (2005). MMP-20 mutation in autosomal

840

Clan MA(M)
M10 | 171. Matrix Metalloproteinase-20/Enamelysin

recessive pigmented hypomaturation amelogenesis imperfecta. J. Med. Genet. 42(3), 271
275. [38] Lee, S.K., Seymen, F., Kang, H.Y., Lee, K.E., Gencay, K., Tuna, B., Kim, J.W. (2010). MMP20 hemopexin domain mutation in amelogenesis imperfecta. J. Dent Res. 89(1), 46
50. [39] Ozdemir, D., Hart, P.S., Ryu, O.H., Choi, S.J., OzdemirKaratas, M., Firatli, E., Piesco, N., Hart, T.C. (2005). MMP20 active-site mutation in hypomaturation amelogenesis imperfecta. J. Dent. Res. 84(11), 1031
1035. [40] Papagerakis, P., Lin, H.K., Lee, K.Y., Hu, Y., Simmer, J.P., Bartlett, J.D., Hu, J.C. (2008). Premature stop codon in MMP20 causing amelogenesis imperfecta. J. Dent. Res. 87(1), 56
59.

[41] Yamada, Y., Yamakoshi, Y., Gerlach, R.F., Hu, C.C., Matsumoto, K., Fukae, M., Oida, S., Bartlett, J.D., Simmer, J.P. (2003). Purification and characterization of enamelysin from secretory stage pig enamel. Arch. Comp. Biol. Tooth Enam. 8, 21
25. [42] Bartlett, J.D., Simmer, J.P. (1999). Proteinases in developing dental enamel. Crit. Rev. Oral Biol. Med. 10(4), 425
441. [43] Simmer, J.P., Hu, J.C. (2002). Expression, structure, and function of enamel proteinases. Connect. Tissue Res. 43(2
3), 441
449. [44] Lu, Y., Papagerakis, P., Yamakoshi, Y., Hu, J.C., Bartlett, J.D., Simmer, J.P. (2008). Functions of KLK4 and MMP-20 in dental enamel formation. Biol. Chem. 389(6), 695
700.

John D. Bartlett The Forsyth Institute, Department of Cytokine Biology, 245 First Street, Cambridge, MA 02142, USA. Email: [email protected] This article is reproduced from the previous edition with revisions made by the Editors. Handbook of Proteolytic Enzymes, 3rd Edn ISBN: 978-0-12-382219-2

© 2013 Elsevier Ltd. All rights reserved. DOI: http://dx.doi.org/10.1016/B978-0-12-382219-2.00171-X