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Binary membranes: A reinterpretation of membrane and myelin structure
Vol. 36, No. 1, 1969
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
BINARY MEMBRANES:
A REINTERPRETATION
OF MEMBRANE
AND MYELIN STRUCTURE F. L. Crane Department Purdue
ReceivedJune
and J. D. Hall
of Biological
University,
Sciences
W. Lafayette,
Indiana
2, 1969
Membrane structure is reinterpreted in terms of a binary Summary: membrane consisting of a double layer of lipoprotein globules. In sectioned and stained mitochondrial preparations the binary membrane appears as a five-layered structure --three dark lines separated by two unstained regions. Electron micrographs published in the literature suggest that other biomembranes may exhibit this binary structure. As a result have
proposed
lipoprotein binary
that
membrane.
two unstained
with
a double fig.
layer
1, both
micrographs
biomembranes structure
are
(2)
(3,4).
This
appearing
binary
rather
can be seen in
than
would
membranes
in
be called
a
plasma
of freeze and stained dark
lines
be consistent
structure
is
micrograph
that
seen
and as
membranes
suggest
membranes.
of
of three
of other
literature
unit
the following
which
we
the yeast
in fixed
electron
examination
in the
for
five-layered
the original
Close
layer should
composed
the lines,
cristae
on the basis
cristae
structure,
in
layer
has been proposed
between
as observed
electron
double
of mitochondrial
membrane
diagrammatically.
this
and Mghlethaler
regions
depicted
(see
model
show a five-layered
with
on mitochondrial
is made up of a double
We feel
A similar
Sections
preparations
in
(1).
by Frey-Wyssling
studies.
studies
the membrane
globules
membrane etch
of fragmentation
and of
other
The five-layer the figures
listed:
list). We feel
enough membrane
that
that it
must
five-layer
structure
be considered
in some cellular
in membranes
as a serious
structures. 174
occurs
alternative
frequently to the unit
Vol. 36, No. 1, 1969
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Figure 1.
A. Section of beef heart mitochondria fixed in osmium of tetroxide and post stained in KMnO4. Arrows mark the three dark lines The membrane thickness as measured from one exterior the binary membrane. stained line to the other exterior line is 90 A. X 710,000.
Arrows mark the diagram of a binary membrane. B. Enlarged Two unstained globular regions are between staining lines.
Since myeI.in
the
derived
of transition layers
is
found
that
schwann
cell
membrane
from
(5)
has a unit
in membrane lost the
during
(7)
myelin.
represents
structure.
myelin
incorporation
the preservation per,ipheral
it
appears structure,
there
It
that
formation. of digitonin
of an additional
layer
Perhaps
this
additional
the original
binary
one of
175
to be binary
appears Napolitano during between layer layers.
three darkly the lines.
while
the
must be some kind one of binary
and Scallen fixation
results
the major
period
or double
minor
(6)
have in
in period
Vol. 36, No. 1, 1969
BIOCHEMICAL AND BIOF’HYSICAL RESEARCH COMMUNICATIONS
In other each
layer
obtained
membranes
would
to 70,000
with
observed
in red blood
and molecular
weights
Fracture etch
procedure
is consistent membrane
with
and Perdue fracture
If then
can be explained lipoprotein packed
to give
surface
consistent The majority coefficients
regions
(13)
membrane
but not
by the
in chloroplasts
freeze
etch
would
planes,
a good
procedure layers
of
be close
e.g.,
globules
by Green is
of two
the globules
uneven
a unit
membrane
composed
on the fracture
with
(14)
as described
of the binary
of a membrane
in the
and Staehelin
of lipoproteins
In some membranes
whereas
to make a
(10,ll).
non-polar
obtained
surfaces
A double
(12).
septum
patterns
on the basis
smooth
(13),
plasma
give
a rough
(15). There
always
are
observed
of one layer changes
between (19)
may not
the coupled could
(16,17)
can induce
In some procedures
may represent
state
to a shift
the binary change in
from
of chloroplasts
structure since
the second 1’76
offers a shift layer.
not
the staining
binary
in membrane
from binary
is
conformational
a shift
differences
and uncoupled
tension
structure
As an alternative,
apparent
conformational
why five-layered
membranes.
be complete.
be related
Certainly
membrane
reasons
in sectioned
For example,
structure.
one layer
two possible
in membranes
structure,
for
the
globules.
membranes
dria
the central
been
from
have 2 S sedimentation
unit
layer
of
have
weights
are also
membranes
by Branton
bilayer
which
be required sizes
central
as described
of a single
(8). plane
50,000
through
a lipid
composed
protein
membrane
of about
of membranes
would
in sectioned
cell
Proteins
a part
of 30 w to 50 A.
size
These
at least
have molecular
diameters
of this
that
(8,9).
amounts
spherical
molecules
50 A globules
freeze
globules
80 A to 100 1 wide.
of proteins
good evidence
in large
with
of protein
membrane
is
be lipoprotein
from membranes
20,000 layer
there
to unit
thickness (18)
to unit
or mitochon-
membrane
mechanical
advantages
or structure
change
in
Vol. 36, No. 1, 1969
BIOCHEMICAL
Pictures
which
show Binary
Schwann cell membrane enclosing axon Schwann cell inner mesaxon around axon at fusion point Bacillus cereus plasma membrane and spore membrane Bacillus subtilis plasma membrane Bacillus mycoides mesosomes Bacillus mycoides plasma membrane inte,stinal microvilli (frog) mitochondrial
cristae
intestinal microvilli axon membrane
mitochondrial
cristae
plasma
membrane
plasma
membrane
liver
cell
(& mitockondria)
endoplasmic
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
reticulum
Membrane
Structure
fig. 6, J. Finean, Circulation 26, 1151 (1963). fig. 1, J. Finean, Circulation 26, 1151 (1963) fig. 17, C. F. Robinow, Circulation 2, 1092 (1963). fig. 15 & 16, C. F. Robinow, Circulation a, 1092 (1963). fig. 8,10,12, C. F. Robinow, Circula;ion-zi 1092 (1963). fin. 7, C. F. Robinow, Circulation 55, io92 (1963). fig. 13, P. W. Brandt, Circulation 26, 1075 (1963). fig. 7 & 8, H. Fernandez Moran, Circulation 26, 1039 (1963). fig. 6, S. Ito, Fed. Proc. 28, 12 (1969). fig. 41b, J. D. Robertson, in Cellular Membranes in Development, Ed. M. Locke, Academic Press, New York, 1964, p. 1. fig. 10 & 11, F. S. Sjostrand in The Membranes, Ed. A. J. Dalton and F. Haguenau, Academic Press, New York, 1968, p. 151. fig. 2, above the arrow, F. S. Sjostrand in The Membranes, Ed. A. J. Dalton and F. Haguenau, Academic Press, New York, 1968, p0 151. fig. 181, D. W. Fawcett, The Cell, Saunders, Philadelphia, 1966, p. 341. Fig. 5, P. Gaylarde and I.. Sarkany, Science 161, 1157 (1968).
ACKNOWLEDGEMENTS Research carried out under grants GM10741 and AM04663 from the National Institutes of Health. F. L. Crane is supported by a Research Career Award K6,21,839 from the National Institute of General Medical Science.
REFERENCES 1.
2. 3.
F. L. Crane, .I. W. Stiles, K. S. Prezbindowski, F. J. Ruzicka, and F,., F. Sun, in Regulatory Functions of Biological Membranes, Ed. J. Jarnefelt, Elsevier, Amsterdam, p. 21 (1968). Ultrastructural Plant Cytology, A. Frey-Wyssling and K. Muhlethaler, Elsevier, Amsterdam, p. 150 (1965). K. S. Prezbindowski, Ph.D. Thesis, Purdue University (1968).
177
Vol. 36, No. 1, 1969
4. 5. 6. 7.
8. 9. 10. 11.
12. 13. 14. 15. 16. 17. 18. 19.
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
K. S. Prezbindowski, F. J. Ruzicka, F. F. Sun, and F. L. Crane, Biochim. Biophys. Res. Commun. 3l, 164 (1968). J. D. Robertson, in Cellular Membranes in Development, Ed. M. Locke, Academic Press, New York, p. 1 (1964). L. M. Napolitano and T. J. Scallen, Anat. Rec. 163, 1 (1969). J. P. Revel and D. W. Hamilton, Anat. Rec. 163, 7 (1969). D. E. Green and J. F. Perdue, Ann. N. Y. Acad. Sci. 137, 667 (1966). E. D. K,qrn, Science 153, 1491 (1966). F. S. Sjostrand, in The Membranes, Ed. A. .I. Dalton and F. Haguenau, Academic Press, New York, p. 151 (1968). E. D. P. DeRobertis, W. W. Nowinski, and F. A. Saez, Cell Biology Philadelphia, p. 113 (1965). Saunders, J. Biol. Chem. 243, 1985 (1968). S. A. Rosenberg and G. Guidotti, D. Branton, Proc. Nat. Acad. Sci. 55, 1048 (1966). L. A. Staehelin, J. Ultrastruct. Res. 22, 326 (1968). D. H. Northcote, Brit. Med. Bull. 24, 107 (1968). C. R. Hackenbrock, J. Cell Biol. 30, 269 (1966). J. T. Pennisfon, R. A. Harris, J. Asai, and D. E. Green, Proc. Nat. Acad. Sci. 59, 624 (1968). S. Izawa and N. E. Good, Plant Physiol. 4l, 544 (1966). D. E. Green, J. Asai, R. A. Harris, and John T. Penniston, Arch. Biochem. Biophys. 125, 684 (1968).
178
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
BINARY MEMBRANES:
A REINTERPRETATION
OF MEMBRANE
AND MYELIN STRUCTURE F. L. Crane Department Purdue
ReceivedJune
and J. D. Hall
of Biological
University,
Sciences
W. Lafayette,
Indiana
2, 1969
Membrane structure is reinterpreted in terms of a binary Summary: membrane consisting of a double layer of lipoprotein globules. In sectioned and stained mitochondrial preparations the binary membrane appears as a five-layered structure --three dark lines separated by two unstained regions. Electron micrographs published in the literature suggest that other biomembranes may exhibit this binary structure. As a result have
proposed
lipoprotein binary
that
membrane.
two unstained
with
a double fig.
layer
1, both
micrographs
biomembranes structure
are
(2)
(3,4).
This
appearing
binary
rather
can be seen in
than
would
membranes
in
be called
a
plasma
of freeze and stained dark
lines
be consistent
structure
is
micrograph
that
seen
and as
membranes
suggest
membranes.
of
of three
of other
literature
unit
the following
which
we
the yeast
in fixed
electron
examination
in the
for
five-layered
the original
Close
layer should
composed
the lines,
cristae
on the basis
cristae
structure,
in
layer
has been proposed
between
as observed
electron
double
of mitochondrial
membrane
diagrammatically.
this
and Mghlethaler
regions
depicted
(see
model
show a five-layered
with
on mitochondrial
is made up of a double
We feel
A similar
Sections
preparations
in
(1).
by Frey-Wyssling
studies.
studies
the membrane
globules
membrane etch
of fragmentation
and of
other
The five-layer the figures
listed:
list). We feel
enough membrane
that
that it
must
five-layer
structure
be considered
in some cellular
in membranes
as a serious
structures. 174
occurs
alternative
frequently to the unit
Vol. 36, No. 1, 1969
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Figure 1.
A. Section of beef heart mitochondria fixed in osmium of tetroxide and post stained in KMnO4. Arrows mark the three dark lines The membrane thickness as measured from one exterior the binary membrane. stained line to the other exterior line is 90 A. X 710,000.
Arrows mark the diagram of a binary membrane. B. Enlarged Two unstained globular regions are between staining lines.
Since myeI.in
the
derived
of transition layers
is
found
that
schwann
cell
membrane
from
(5)
has a unit
in membrane lost the
during
(7)
myelin.
represents
structure.
myelin
incorporation
the preservation per,ipheral
it
appears structure,
there
It
that
formation. of digitonin
of an additional
layer
Perhaps
this
additional
the original
binary
one of
175
to be binary
appears Napolitano during between layer layers.
three darkly the lines.
while
the
must be some kind one of binary
and Scallen fixation
results
the major
period
or double
minor
(6)
have in
in period
Vol. 36, No. 1, 1969
BIOCHEMICAL AND BIOF’HYSICAL RESEARCH COMMUNICATIONS
In other each
layer
obtained
membranes
would
to 70,000
with
observed
in red blood
and molecular
weights
Fracture etch
procedure
is consistent membrane
with
and Perdue fracture
If then
can be explained lipoprotein packed
to give
surface
consistent The majority coefficients
regions
(13)
membrane
but not
by the
in chloroplasts
freeze
etch
would
planes,
a good
procedure layers
of
be close
e.g.,
globules
by Green is
of two
the globules
uneven
a unit
membrane
composed
on the fracture
with
(14)
as described
of the binary
of a membrane
in the
and Staehelin
of lipoproteins
In some membranes
whereas
to make a
(10,ll).
non-polar
obtained
surfaces
A double
(12).
septum
patterns
on the basis
smooth
(13),
plasma
give
a rough
(15). There
always
are
observed
of one layer changes
between (19)
may not
the coupled could
(16,17)
can induce
In some procedures
may represent
state
to a shift
the binary change in
from
of chloroplasts
structure since
the second 1’76
offers a shift layer.
not
the staining
binary
in membrane
from binary
is
conformational
a shift
differences
and uncoupled
tension
structure
As an alternative,
apparent
conformational
why five-layered
membranes.
be complete.
be related
Certainly
membrane
reasons
in sectioned
For example,
structure.
one layer
two possible
in membranes
structure,
for
the
globules.
membranes
dria
the central
been
from
have 2 S sedimentation
unit
layer
of
have
weights
are also
membranes
by Branton
bilayer
which
be required sizes
central
as described
of a single
(8). plane
50,000
through
a lipid
composed
protein
membrane
of about
of membranes
would
in sectioned
cell
Proteins
a part
of 30 w to 50 A.
size
These
at least
have molecular
diameters
of this
that
(8,9).
amounts
spherical
molecules
50 A globules
freeze
globules
80 A to 100 1 wide.
of proteins
good evidence
in large
with
of protein
membrane
is
be lipoprotein
from membranes
20,000 layer
there
to unit
thickness (18)
to unit
or mitochon-
membrane
mechanical
advantages
or structure
change
in
Vol. 36, No. 1, 1969
BIOCHEMICAL
Pictures
which
show Binary
Schwann cell membrane enclosing axon Schwann cell inner mesaxon around axon at fusion point Bacillus cereus plasma membrane and spore membrane Bacillus subtilis plasma membrane Bacillus mycoides mesosomes Bacillus mycoides plasma membrane inte,stinal microvilli (frog) mitochondrial
cristae
intestinal microvilli axon membrane
mitochondrial
cristae
plasma
membrane
plasma
membrane
liver
cell
(& mitockondria)
endoplasmic
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
reticulum
Membrane
Structure
fig. 6, J. Finean, Circulation 26, 1151 (1963). fig. 1, J. Finean, Circulation 26, 1151 (1963) fig. 17, C. F. Robinow, Circulation 2, 1092 (1963). fig. 15 & 16, C. F. Robinow, Circulation a, 1092 (1963). fig. 8,10,12, C. F. Robinow, Circula;ion-zi 1092 (1963). fin. 7, C. F. Robinow, Circulation 55, io92 (1963). fig. 13, P. W. Brandt, Circulation 26, 1075 (1963). fig. 7 & 8, H. Fernandez Moran, Circulation 26, 1039 (1963). fig. 6, S. Ito, Fed. Proc. 28, 12 (1969). fig. 41b, J. D. Robertson, in Cellular Membranes in Development, Ed. M. Locke, Academic Press, New York, 1964, p. 1. fig. 10 & 11, F. S. Sjostrand in The Membranes, Ed. A. J. Dalton and F. Haguenau, Academic Press, New York, 1968, p. 151. fig. 2, above the arrow, F. S. Sjostrand in The Membranes, Ed. A. J. Dalton and F. Haguenau, Academic Press, New York, 1968, p0 151. fig. 181, D. W. Fawcett, The Cell, Saunders, Philadelphia, 1966, p. 341. Fig. 5, P. Gaylarde and I.. Sarkany, Science 161, 1157 (1968).
ACKNOWLEDGEMENTS Research carried out under grants GM10741 and AM04663 from the National Institutes of Health. F. L. Crane is supported by a Research Career Award K6,21,839 from the National Institute of General Medical Science.
REFERENCES 1.
2. 3.
F. L. Crane, .I. W. Stiles, K. S. Prezbindowski, F. J. Ruzicka, and F,., F. Sun, in Regulatory Functions of Biological Membranes, Ed. J. Jarnefelt, Elsevier, Amsterdam, p. 21 (1968). Ultrastructural Plant Cytology, A. Frey-Wyssling and K. Muhlethaler, Elsevier, Amsterdam, p. 150 (1965). K. S. Prezbindowski, Ph.D. Thesis, Purdue University (1968).
177
Vol. 36, No. 1, 1969
4. 5. 6. 7.
8. 9. 10. 11.
12. 13. 14. 15. 16. 17. 18. 19.
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
K. S. Prezbindowski, F. J. Ruzicka, F. F. Sun, and F. L. Crane, Biochim. Biophys. Res. Commun. 3l, 164 (1968). J. D. Robertson, in Cellular Membranes in Development, Ed. M. Locke, Academic Press, New York, p. 1 (1964). L. M. Napolitano and T. J. Scallen, Anat. Rec. 163, 1 (1969). J. P. Revel and D. W. Hamilton, Anat. Rec. 163, 7 (1969). D. E. Green and J. F. Perdue, Ann. N. Y. Acad. Sci. 137, 667 (1966). E. D. K,qrn, Science 153, 1491 (1966). F. S. Sjostrand, in The Membranes, Ed. A. .I. Dalton and F. Haguenau, Academic Press, New York, p. 151 (1968). E. D. P. DeRobertis, W. W. Nowinski, and F. A. Saez, Cell Biology Philadelphia, p. 113 (1965). Saunders, J. Biol. Chem. 243, 1985 (1968). S. A. Rosenberg and G. Guidotti, D. Branton, Proc. Nat. Acad. Sci. 55, 1048 (1966). L. A. Staehelin, J. Ultrastruct. Res. 22, 326 (1968). D. H. Northcote, Brit. Med. Bull. 24, 107 (1968). C. R. Hackenbrock, J. Cell Biol. 30, 269 (1966). J. T. Pennisfon, R. A. Harris, J. Asai, and D. E. Green, Proc. Nat. Acad. Sci. 59, 624 (1968). S. Izawa and N. E. Good, Plant Physiol. 4l, 544 (1966). D. E. Green, J. Asai, R. A. Harris, and John T. Penniston, Arch. Biochem. Biophys. 125, 684 (1968).
178