Effect of various glycosidase treatments on the resistance of the HIV‐1 envelope to degradation

FEBS Letters

Papandreou, Marie-Jeanne¹  Fenouillet, Emmanuel¹ 

[1] CNRS, UMR 6560, IFR Jean Roche, Faculté de Médecine Nord, Marseille, France

关键词: Conformation;    Denaturation;    Envelope;    Glycosylation;    HIV;    Proteolysis;    ABB;    ammonium bicarbonate buffer;    BHK;    baby hamster kidney;    CD;    cluster of differentiation;    CHO;    chinese hamster ovary;    CHO+;    glycosylated molecule;    CHO−;    N-glycanase-treated molecule;    EH;    endo-β-N-acetylglucosaminidase H;    gp160;    envelope precursor glycoprotein;    gp120;    outer membrane envelope glycoprotein;    gp41;    transmembrane envelope glycoprotein;    HIV-1;    human immunodeficiency virus type 1;    HMT;    high-mannose-type structures;    HMT−;    endo-β-N-acetylglucosaminidase H-treated molecule;    HPLC;    high-pressure liquid chromatography;    NG;    N-glycanase;    PBS;    phosphate-buffered saline;    PBSC;    PBS 2% casein;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecyl sulfate;    Sial;    sialidase;    Sial−;    sialidase-treated molecule;    V3;    third variable domain of the outer membrane envelope glycoprotein;   

DOI  :  10.1016/S0014-5793(97)00273-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using a CD4-binding assay to assess the conformation of the human immunodeficiency virus envelope glycoprotein (CHO⁺Env), we studied the effect of treatment with various glycosidases on the stability of Env in denaturing environments and in biological media: cleavage from Env of either high-mannose-type glycans (HMT⁻Env) by endoglycosidase H or sialic acid residues (Sial⁻Env) by sialidase did not alter Env stability whereas its complete deglycosylation (CHO⁻Env) by N-glycanase had a large effect. The influence of glycan removal on Env sensitivity to proteases was also studied. Thrombin cleavage within V3 was affected by N-glycanase treatment; both HMT⁻Env and CHO⁻Env displayed an increased sensitivity to other endoproteases. Thus, partial deglycosylation increases Env sensitivity to proteases but only its total deglycosylation alters its stability.

【 授权许可】

Unknown