The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments

Mikyung Kim, Bing Chen, Rebecca E. Hussey, Yasmin Chishti, David Montefiori, James A. Hoxie, Olwyn Byron, Gordon Campbell, Stephen C. Harrison, Ellis L. Reinherz

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Human and simian immunodeficiency viruses infect host lymphoid cells by binding CD4 molecules via their gp160 envelope glycoproteins. Biochemical studies on recombinant SIVmac32H (pJ5) envelope ectodomain gp140 precursor protein show that the envelope is a trimer. Using size exclusion chromatography, quantitative amino acid analysis, analytical ultracentrifugation, and CD4-based competition assay, we demonstrate that the stoichiometry of CD4 receptor-oligomeric envelope interaction is 1:1. By contrast, Fab fragments of both neutralizing and non-neutralizing monoclonal antibodies bind at a 3:1 ratio. Thus, despite displaying equivalent CD4 binding sites on each of the three gp140 protomers within an uncleaved trimer, only one site binds the soluble 4-domain human CD4 extracellular segment. The anti-cooperativity and the faster koff of gp140 trimer:CD4 versus gp120 monomer:CD4 interaction suggest that CD4-induced conformational change is impeded in the intact envelope. The implications of these findings for immunity against human immunodeficiency virus and simian immunodeficiency virus are discussed.

Original languageEnglish (US)
Pages (from-to)42667-42676
Number of pages10
JournalJournal of Biological Chemistry
Volume276
Issue number46
DOIs
StatePublished - Nov 16 2001
Externally publishedYes

Fingerprint

Immunoglobulin Fragments
Immunoglobulin Fab Fragments
Viruses
Stoichiometry
Anti-Idiotypic Antibodies
Simian Immunodeficiency Virus
CD4 Antigens
Glycoproteins
Antibodies
HIV
Protein Precursors
Size exclusion chromatography
Ultracentrifugation
Protein Subunits
Gel Chromatography
Immunity
Assays
Monomers
Binding Sites
Monoclonal Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments. / Kim, Mikyung; Chen, Bing; Hussey, Rebecca E.; Chishti, Yasmin; Montefiori, David; Hoxie, James A.; Byron, Olwyn; Campbell, Gordon; Harrison, Stephen C.; Reinherz, Ellis L.

In: Journal of Biological Chemistry, Vol. 276, No. 46, 16.11.2001, p. 42667-42676.

Research output: Contribution to journalArticle

Kim, M, Chen, B, Hussey, RE, Chishti, Y, Montefiori, D, Hoxie, JA, Byron, O, Campbell, G, Harrison, SC & Reinherz, EL 2001, 'The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments', Journal of Biological Chemistry, vol. 276, no. 46, pp. 42667-42676. https://doi.org/10.1074/jbc.M104166200
Kim, Mikyung ; Chen, Bing ; Hussey, Rebecca E. ; Chishti, Yasmin ; Montefiori, David ; Hoxie, James A. ; Byron, Olwyn ; Campbell, Gordon ; Harrison, Stephen C. ; Reinherz, Ellis L. / The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 46. pp. 42667-42676.
@article{9fbaf3f33afa405bbbfec8f0e700c6a9,
title = "The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments",
abstract = "Human and simian immunodeficiency viruses infect host lymphoid cells by binding CD4 molecules via their gp160 envelope glycoproteins. Biochemical studies on recombinant SIVmac32H (pJ5) envelope ectodomain gp140 precursor protein show that the envelope is a trimer. Using size exclusion chromatography, quantitative amino acid analysis, analytical ultracentrifugation, and CD4-based competition assay, we demonstrate that the stoichiometry of CD4 receptor-oligomeric envelope interaction is 1:1. By contrast, Fab fragments of both neutralizing and non-neutralizing monoclonal antibodies bind at a 3:1 ratio. Thus, despite displaying equivalent CD4 binding sites on each of the three gp140 protomers within an uncleaved trimer, only one site binds the soluble 4-domain human CD4 extracellular segment. The anti-cooperativity and the faster koff of gp140 trimer:CD4 versus gp120 monomer:CD4 interaction suggest that CD4-induced conformational change is impeded in the intact envelope. The implications of these findings for immunity against human immunodeficiency virus and simian immunodeficiency virus are discussed.",
author = "Mikyung Kim and Bing Chen and Hussey, {Rebecca E.} and Yasmin Chishti and David Montefiori and Hoxie, {James A.} and Olwyn Byron and Gordon Campbell and Harrison, {Stephen C.} and Reinherz, {Ellis L.}",
year = "2001",
month = "11",
day = "16",
doi = "10.1074/jbc.M104166200",
language = "English (US)",
volume = "276",
pages = "42667--42676",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "46",

}

TY - JOUR

T1 - The Stoichiometry of Trimeric SIV Glycoprotein Interaction with CD4 Differs from That of Anti-envelope Antibody Fab Fragments

AU - Kim, Mikyung

AU - Chen, Bing

AU - Hussey, Rebecca E.

AU - Chishti, Yasmin

AU - Montefiori, David

AU - Hoxie, James A.

AU - Byron, Olwyn

AU - Campbell, Gordon

AU - Harrison, Stephen C.

AU - Reinherz, Ellis L.

PY - 2001/11/16

Y1 - 2001/11/16

N2 - Human and simian immunodeficiency viruses infect host lymphoid cells by binding CD4 molecules via their gp160 envelope glycoproteins. Biochemical studies on recombinant SIVmac32H (pJ5) envelope ectodomain gp140 precursor protein show that the envelope is a trimer. Using size exclusion chromatography, quantitative amino acid analysis, analytical ultracentrifugation, and CD4-based competition assay, we demonstrate that the stoichiometry of CD4 receptor-oligomeric envelope interaction is 1:1. By contrast, Fab fragments of both neutralizing and non-neutralizing monoclonal antibodies bind at a 3:1 ratio. Thus, despite displaying equivalent CD4 binding sites on each of the three gp140 protomers within an uncleaved trimer, only one site binds the soluble 4-domain human CD4 extracellular segment. The anti-cooperativity and the faster koff of gp140 trimer:CD4 versus gp120 monomer:CD4 interaction suggest that CD4-induced conformational change is impeded in the intact envelope. The implications of these findings for immunity against human immunodeficiency virus and simian immunodeficiency virus are discussed.

AB - Human and simian immunodeficiency viruses infect host lymphoid cells by binding CD4 molecules via their gp160 envelope glycoproteins. Biochemical studies on recombinant SIVmac32H (pJ5) envelope ectodomain gp140 precursor protein show that the envelope is a trimer. Using size exclusion chromatography, quantitative amino acid analysis, analytical ultracentrifugation, and CD4-based competition assay, we demonstrate that the stoichiometry of CD4 receptor-oligomeric envelope interaction is 1:1. By contrast, Fab fragments of both neutralizing and non-neutralizing monoclonal antibodies bind at a 3:1 ratio. Thus, despite displaying equivalent CD4 binding sites on each of the three gp140 protomers within an uncleaved trimer, only one site binds the soluble 4-domain human CD4 extracellular segment. The anti-cooperativity and the faster koff of gp140 trimer:CD4 versus gp120 monomer:CD4 interaction suggest that CD4-induced conformational change is impeded in the intact envelope. The implications of these findings for immunity against human immunodeficiency virus and simian immunodeficiency virus are discussed.

UR - http://www.scopus.com/inward/record.url?scp=0035900737&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035900737&partnerID=8YFLogxK

U2 - 10.1074/jbc.M104166200

DO - 10.1074/jbc.M104166200

M3 - Article

C2 - 11544255

AN - SCOPUS:0035900737

VL - 276

SP - 42667

EP - 42676

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 46

ER -