Identification of the U‐937 membrane‐associated proteinase interacting with the V3 loop of HIV‐1 gp12O as cathepsin G

FEBS Letters

Di Martino-Ferrer, Michèle²  Gauthier, Francis²  Pignede, Georges¹  Avril, Laurence-Emmanuelle²  Séman, Michel¹ 

[1] Laboratoire d'Immunodifferenciation, University Denis Diderot, Paris France;Laboratoire d'Enzymologie et Chimie des Protéines, Centre National de la Recherche Scientifique URA 1334, University François Rabelais, Faculty of Medicine, 2 bis Boulevard Tonnellé, F 37032 Tours Cedex, France

关键词: HIV-1;    gp12O V3 loop;    Proteinase;    Cathepsin G;    ABTS;    2;    2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid);    DTNB;    (5;    5′-dithiobis-(2-nitrobenzoic acid);    NaCl/Pi;    phosphate-buffered saline;    PCR;    polymerase chain reaction;    STI;    soybean trypsin inhibitor;    Suc-Ala-Ala-Pro-Phe-SBzl;    N-α-Cbz-alanyl-alanyl-prolyl-phenylalanyl-thiobenzylester·HCl;    TFA;    trifluoroacetic acid;    Z-Lys-SBzl;    N-α-CbZ-l-Lysine-thiobenzylester·HCl;   

DOI  :  10.1016/0014-5793(94)00410-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have purified a serine proteinase from the membrane of U-937 cells that was inhibited in a tight-binding manner by recombinant gp120 and by peptides mimicking the V3 loop of gp12O [(1993) FEBS Lett. 317, 167–172]. This proteinase has now been characterized, both structurally and functionally. It has a dual trypsin- and chymotrypsin-like specificity, and N-terminal sequence analysis of the first 32 residues indicates complete identity with leukocyte cathepsin G. Cathepsin G-like material was located at the surface of U-937 cells using a monoclonal antibody directed against leukocyte cathepsin G, and polyclonal anti-cathepsin G antibodies precipitated the purified proteinase. However, the U-937 enzyme differs slightly from commercial leukocyte cathepsin G in its apparent M _r because of different glycosylation. No other protein structurally related to cathepsin G was found upon screening a U-937 cDNA library using several oligonucleotide probes constructed from the membrane proteinase N-terminal amino acid sequence. The possible interaction of a cathepsin G-like proteinase at the surface of U-937 cells with the V3 loop of HIV-1 gp120 is discussed.

【 授权许可】

Unknown