Thiol/disulfide exchange is a prerequisite for CXCR4-tropic HIV-1 envelope-mediated T-cell fusion during viral entry.
Attachment of gp120 to CD4 during HIV-1 entry triggers structural rearrangement in gp120 that enables binding to an appropriate coreceptor. Following coreceptor engagement, additional conformational changes occur in the envelope (Env), resulting in fusion of virion and cell membranes. Catalysts with redox-isomerase activity, such as protein disulfide isomerase (PDI), facilitate Env conversion from its inactive to its fusion-competent conformation. We report here that anti-PDI agents effectively block CXCR4 Env-mediated fusion and spread of virus infection. Exogenously added PDI, in turn, can rescue fusion from this blockade. We further find that PDI facilitates thiol/disulfide rearrangement in gp120 during conformational change, whereas inhibition of this redox shuffling prevents gp41 from assuming the fusogenic 6-helix bundle conformation. At the virus-cell contact site, gp120 induces assembly of PDI, CD4, and CXCR4 into a tetramolecular protein complex serving as a portal for viral entry. Our findings support the hypothesis that Env conformational change depends on a well-coordinated action of a tripartite system in which PDI works in concert with the receptor and the coreceptor to effectively lower the activation energy barrier required for Env conformational rearrangement.
Center for Drug Evaluation and Research, Food and Drug Adminiatration, Bethesda, MD, USA. markovic@cber.fda.gov
Ingrid
I
Markovic
Tzanko S
TS
Stantchev
Karen H
KH
Fields
Linda J
LJ
Tiffany
Melanija
M
Tomiç
Carol D
CD
Weiss
Christopher C
CC
Broder
Klaus
K
Strebel
Kathleen A
KA
Clouse
D000818:Animals; D001665:Binding Sites; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002459:Cell Fusion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D004220:Disulfides; D004306:Dose-Response Relationship, Immunologic; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry; D018613:Microscopy, Confocal; D010084:Oxidation-Reduction; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011817:Rabbits; D019718:Receptors, CXCR4 / Q000378:metabolism*; D013601:T-Lymphocytes / Q000821:virology*; D013997:Time Factors
D004220:Disulfides; D004791:Enzyme Inhibitors; D015699:HIV Envelope Protein gp120; D019718:Receptors, CXCR4; D019704:Protein Disulfide-Isomerases; D002118:Calcium