Disulfide bonds stabilize JC virus capsid-like structure by protecting calcium ions from chelation.
To investigate the role of disulfide bonds in the capsid structure, a recombinant JC virus-like particle (VLP) was used. The major capsid protein, VP1, of the JC virus was expressed in yeast cells. The yeast-expressed VP1 was self-assembled into a VLP. Disulfide bonds were found in the VLP which caused dimeric and trimeric VP1 linkages as demonstrated by non-reducing SDS-PAGE. The VLP remained intact when disulfide bonds were reduced by dithiothreitol. The VLP without disulfide bonds could be disassembled into capsomeres by EGTA alone, but those with disulfide bonds could not be disassembled by EGTA. Capsomeres were reassembled into VLPs in the presence of calcium ions. Capsomeres formed irregular aggregations instead of VLPs when treated with diamide to reconstitute the disulfide bonds. These results indicate that disulfide bonds play an important role in maintaining the integrity of the JC VLP by protecting calcium ions from chelation.
Chen
P L
PL
Institute of Molecular Biology, National Chung Cheng University, Taiwan, ROC.
Wang
M
M
Ou
W C
WC
Lii
C K
CK
Chen
L S
LS
Chang
D
D
D002118:Calcium / Q000378:metabolism*; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D036022:Capsid Proteins; D002614:Chelating Agents / Q000737:chemistry / Q000378:metabolism*; D003958:Diamide / Q000737:chemistry / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D004533:Egtazic Acid / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007577:JC Virus / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D019163:Reducing Agents / Q000737:chemistry / Q000494:pharmacology; D012441:Saccharomyces cerevisiae; D014162:Transfection; D014771:Virion / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism
D036022:Capsid Proteins; D002614:Chelating Agents; D004220:Disulfides; D019163:Reducing Agents; C062481:VP1 protein, polyomavirus; D003958:Diamide; D004533:Egtazic Acid; D002118:Calcium