TY - JOUR
T1 - Three-dimensional organization of retroviral capsid proteins on a lipid monolayer
AU - McDermott, Jason
AU - Mayo, Keith
AU - Barklis, Eric
N1 - Funding Information:
We are grateful to Mike Schmid and Stephen Fuller for continual image processing help and advice; and to Elizabeth Kubalek-Wilson for the nickel-chelating lipid, DOGS. Sonya Karanjia contributed invaluable assistance in protein purification and crystallization; Robin Barklis performed tilt axis determinations; and Doug Huseby and Guy Zuber provided helpful advice. This work was supported by NIH grants GM52914 and GM60170 to E.B., and by NIH pre-doctoral training support (A107472) to J.M.
PY - 2000/9/8
Y1 - 2000/9/8
N2 - We have used a method for the two-dimensional crystallization of retro-viral structural proteins to obtain a three-dimensional structure of negatively stained, membrane-bound, histidine-tagged Moloney murine leukemia virus (M-MuLV) capsid protein (his-MoCA) arrays. Tilted and untilted micrographs from crystals formed by purified his-MoCA proteins incubated beneath lipid monolayers containing nickel-chelating lipids were used in 3D reconstructions. The 2D crystals had unit cell dimensions of a = 72.6 Å, b = 72.5 Å and γ = 119.5°, but appeared to have no intrinsic symmetry (p1) in 3D, in contrast to the trigonal or hexagonal appearance of their 2D projections. Membrane-bound his-MoCA proteins showed a strand-like organization, apparently with dimer building blocks, Membrane-proximal regions, or putative N-terminal domains (NTDs), dimerized with different partners than the membrane-distal putative C-terminal domains (CTDs). Evidence also suggests that CTDs can adopt alternate orientations relative to their NTDs, forming inter-strand connections, Our results are consistent with helical-spiral models for retrovirus particle assembly, but are not easily reconcilable with icosa- hedral models. (C) 2000 Academic Press.
AB - We have used a method for the two-dimensional crystallization of retro-viral structural proteins to obtain a three-dimensional structure of negatively stained, membrane-bound, histidine-tagged Moloney murine leukemia virus (M-MuLV) capsid protein (his-MoCA) arrays. Tilted and untilted micrographs from crystals formed by purified his-MoCA proteins incubated beneath lipid monolayers containing nickel-chelating lipids were used in 3D reconstructions. The 2D crystals had unit cell dimensions of a = 72.6 Å, b = 72.5 Å and γ = 119.5°, but appeared to have no intrinsic symmetry (p1) in 3D, in contrast to the trigonal or hexagonal appearance of their 2D projections. Membrane-bound his-MoCA proteins showed a strand-like organization, apparently with dimer building blocks, Membrane-proximal regions, or putative N-terminal domains (NTDs), dimerized with different partners than the membrane-distal putative C-terminal domains (CTDs). Evidence also suggests that CTDs can adopt alternate orientations relative to their NTDs, forming inter-strand connections, Our results are consistent with helical-spiral models for retrovirus particle assembly, but are not easily reconcilable with icosa- hedral models. (C) 2000 Academic Press.
KW - Capsid
KW - Electron microscopy
KW - Gag
KW - Membrane
KW - Retrovirus
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U2 - 10.1006/jmbi.2000.4030
DO - 10.1006/jmbi.2000.4030
M3 - Article
C2 - 10964565
AN - SCOPUS:0034622934
SN - 0022-2836
VL - 302
SP - 121
EP - 133
JO - Journal of molecular biology
JF - Journal of molecular biology
IS - 1
ER -