Role of N-linked Glycans in the Interaction Between the Envelope Glycoprotein of Human Immunodeficiency Virus and Its CD4 Cellular Receptor. Structural Enzymatic Analysis

Overview

Journal J Exp Med

Specialties Allergy & Immunology
General Medicine

Date 1989 Mar 1

PMID 2538547

Citations 29

Authors

E Fenouillet

J C Gluckman

D Guetard

L Montagnier

E Bahraoui

Affiliations

Soon will be listed here.

Abstract

gp120 and CD4 are two glycoproteins that are considered to interact together to allow the binding of HIV to CD4+ cells. We have utilized enzymatic digestion by endoglycosidases in order to analyze N-linked carbohydrate chains of these proteins and their possible role in the interaction of gp120 or gp160 with CD4. SDS denaturation was not necessary to obtain optimal deglycosylation of either molecule, but deglycosylation of CD4, nonetheless, depended on the presence of 1% Triton X-100. Endo H and Endo F that cleave high mannose type and biantennary glycans diminish the molecular mass of the glycoproteins from 120 or 160 Kd to 90 or 130 Kd, respectively; but these enzymes had no action on CD4 glycans. Endo F N-glycanase mixture, which acts on all glycan species, including triantennary chains, led to complete deglycosylation of gp120/160 and of CD4. Therefore, probably half of the glycan moieties of gp120/160 are composed of high mannose and biantennary chains, the other half being triantennary species. The carbohydrate structures of CD4 seems to be triantennary chains. To analyze the binding of gp120/160 to CD4, we used a molecular assay in which an mAb (110-4) coupled to Sepharose CL4B allowed the attachment of soluble gp120/160 to the beads; 125I-sCD4 was then added to measure the binding of CD4 to different amounts of gp120/160. Binding to gp160 was not modified when using completely deglycosylated 125I-sCD4, while deglycosylation of gp120 or of gp160 resulted in the decrease of the binding to native CD4 by two- and fivefold, respectively. Native and deglycosylated gp120/160 bound to CD4+ cells with comparable affinities. In addition, deglycosylated gp120 displaced 125I-gp160 binding to CD4+ cells and inhibited fusion of fresh Molt-T4 cells with CEM HIV1- or HIV2-infected cells to the same extent. Taken together, these results indicate that carbohydrates of CD4 and of gp120/160 do not play a significant role in the in vitro interaction between these two molecules.

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References

Nakamura K, Compans R . Effects of glucosamine, 2-deoxyglucose, and tunicamycin on glycosylation, sulfation, and assembly of influenza viral proteins. Virology. 1978; 84(2):303-19. DOI: 10.1016/0042-6822(78)90250-7. View

Duksin D, Bornstein P . Changes in surface properties of normal and transformed cells caused by tunicamycin, an inhibitor of protein glycosylation. Proc Natl Acad Sci U S A. 1977; 74(8):3433-7. PMC: 431594. DOI: 10.1073/pnas.74.8.3433. View

Muller R . Calculation of average antibody affinity in anti-hapten sera from data obtained by competitive radioimmunoassay. J Immunol Methods. 1980; 34(4):345-52. DOI: 10.1016/0022-1759(80)90107-6. View

Gibson R, Kornfeld S, Schlesinger S . The effect of oligosaccharide chains of different sizes on the maturation and physical properties of the G protein of vesicular stomatitis virus. J Biol Chem. 1981; 256(1):456-62. View

Gitelman A, Berezin V, Kharitonenkov I . The role of carbohydrate in determining the immunochemical properties of the hemagglutinin of influenza A virus. Arch Virol. 1981; 67(3):253-66. DOI: 10.1007/BF01318135. View

Crimmins D, Schlesinger S . Physical properties of the glycoprotein of vesicular stomatitis virus measured by intrinsic fluorescence and aggregation. Biochemistry. 1982; 21(14):3518-24. DOI: 10.1021/bi00257a040. View

Elder J, Alexander S . endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982; 79(15):4540-4. PMC: 346710. DOI: 10.1073/pnas.79.15.4540. View

Lennarz W . Glycoprotein synthesis and embryonic development. CRC Crit Rev Biochem. 1983; 14(4):257-72. DOI: 10.3109/10409238309102795. View

Klatzmann D, Barre-Sinoussi F, Nugeyre M, Danquet C, Vilmer E, Griscelli C . Selective tropism of lymphadenopathy associated virus (LAV) for helper-inducer T lymphocytes. Science. 1984; 225(4657):59-63. DOI: 10.1126/science.6328660. View

10.

PLUMMER Jr T, Elder J, Alexander S, Phelan A, Tarentino A . Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations. J Biol Chem. 1984; 259(17):10700-4. View

11.

Klatzmann D, Champagne E, Chamaret S, Gruest J, Guetard D, Hercend T . T-lymphocyte T4 molecule behaves as the receptor for human retrovirus LAV. Nature. 1984; 312(5996):767-8. DOI: 10.1038/312767a0. View

12.

McDougal J, Cort S, Kennedy M, Cabridilla C, Feorino P, Francis D . Immunoassay for the detection and quantitation of infectious human retrovirus, lymphadenopathy-associated virus (LAV). J Immunol Methods. 1985; 76(1):171-83. DOI: 10.1016/0022-1759(85)90489-2. View

13.

Edelman G . Cell adhesion and the molecular processes of morphogenesis. Annu Rev Biochem. 1985; 54:135-69. DOI: 10.1146/annurev.bi.54.070185.001031. View

14.

KORNFELD R, Kornfeld S . Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985; 54:631-64. DOI: 10.1146/annurev.bi.54.070185.003215. View

15.

Veronese F, Devico A, Copeland T, Oroszlan S, Gallo R, Sarngadharan M . Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science. 1985; 229(4720):1402-5. DOI: 10.1126/science.2994223. View

16.

Tarentino A, Gomez C, PLUMMER Jr T . Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F. Biochemistry. 1985; 24(17):4665-71. DOI: 10.1021/bi00338a028. View

17.

von Figura K, Hasilik A . Lysosomal enzymes and their receptors. Annu Rev Biochem. 1986; 55:167-93. DOI: 10.1146/annurev.bi.55.070186.001123. View

18.

McDougal J, Nicholson J, Cross G, Cort S, Kennedy M, Mawle A . Binding of the human retrovirus HTLV-III/LAV/ARV/HIV to the CD4 (T4) molecule: conformation dependence, epitope mapping, antibody inhibition, and potential for idiotypic mimicry. J Immunol. 1986; 137(9):2937-44. View

19.

Desai S, Kalyanaraman V, Casey J, Srinivasan A, Andersen P, Devare S . Molecular cloning and primary nucleotide sequence analysis of a distinct human immunodeficiency virus isolate reveal significant divergence in its genomic sequences. Proc Natl Acad Sci U S A. 1986; 83(21):8380-4. PMC: 386932. DOI: 10.1073/pnas.83.21.8380. View

20.

Lifson J, Coutre S, Huang E, Engleman E . Role of envelope glycoprotein carbohydrate in human immunodeficiency virus (HIV) infectivity and virus-induced cell fusion. J Exp Med. 1986; 164(6):2101-6. PMC: 2188479. DOI: 10.1084/jem.164.6.2101. View