Inhibition of U-937 membrane-associated cathepsin G by GP120 (IIIB) and V3 loop-derived peptides from several strains of HIV-1.
A cell surface-associated cathepsin G has been reported to be a possible complementary factor for HIV-1 infection of U-937 cells. The effect of recombinant gp120 (IIIB) and a series of V3 loop peptides derived from the sequence of different strains of HIV on the activity of U-937 cathepsin G was assayed. The sequence on the N-terminal side of the highly conserved GPGRAF V3 loop segment was required for interaction with cathepsin G. The inhibition was stable for several hours and there was no cleavage of the peptides derived from the HIV-1(IIIB) strain. Recombinant gp120 (IIIB) also remained uncleaved after incubation with cathepsin G for 3 h, but some cleavage occurred, generating 2 fragments (50 kDa and 70 kDa), after 16 h. Linear peptides derived from HIV-1 Mal, ELI, MN, CDC4 and SF162 strains, and consensus V3 peptides all had inhibitory properties towards cathepsin G, although they were significantly cleaved after one hour. The cleavage site was at the carboxy-terminus of Tyr323 which is conserved in all these HIV-1 strains but not in HIV-1(IIIB). There was no cleavage at the Arg residue of the GPGRAF sequence, whatever the V3 peptide sequence, the amount of proteinase, or the incubation time. We conclude that the inhibition of membrane-associated cathepsin G of U-937 cells by the gp120 V3 loop of HIV-1 does not occur via a Kunitz-type mechanism, and that the proteinase-V3 loop interaction does not result in a significant cleavage of the V3 loop, though it has been suggested that this event is required for the entry phase of the virus.
Avril
L E
LE
Laboratory of Enzymology and Protein Chemistry, CNRS URA 1334, University François Rabelais, Tours, France.
di Martino-Ferrer
M
M
Brillard-Bourdet
M
M
Gauthier
F
F
D000595:Amino Acid Sequence; D056649:Cathepsin G; D002403:Cathepsins / Q000037:antagonists & inhibitors*; D016384:Consensus Sequence; D015699:HIV Envelope Protein gp120 / Q000378:metabolism / Q000494:pharmacology*; D015497:HIV-1 / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D011485:Protein Binding; D012697:Serine Endopeptidases; D013329:Structure-Activity Relationship
D015699:HIV Envelope Protein gp120; D009842:Oligopeptides; D002403:Cathepsins; D012697:Serine Endopeptidases; D056649:Cathepsin G