Sub-site preferences of the aspartic proteinase from the human immunodeficiency virus, HIV-1.
A series of synthetic, chromogenic substrates for HIV-1 proteinase with the general structure Ala-Thr-His-Xaa-Yaa-Zaa*Nph-Val-Arg-Lys-Ala was synthesised with a variety of residues introduced into the Xaa, Yaa and Zaa positions. Kinetics parameters for hydrolysis of each peptide by HIV-1 proteinase at pH 4.7, 37 degrees C and u = 1.0 M were measured spectrophotometrically and/or by reverse phase FPLC. A variety of residues was found to be acceptable in the P3 position whilst hydrophobic/aromatic residues were preferable in P1. The nature of the residue occupying the P2 position had a strong influence on kcat (with little effect on Km); beta-branched residues Val or Ile in this position resulted in considerably faster peptide hydrolysis than when e.g. the Leu-containing analogue was present in P2.
Konvalinka
J
J
Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Science, Praha.
Strop
P
P
Velek
J
J
Cerna
V
V
Kostka
V
V
Phylip
L H
LH
Richards
A D
AD
Dunn
B M
BM
Kay
J
J
D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000378:metabolism*; D015684:Gene Products, pol / Q000378:metabolism*; D016333:HIV Protease; D015497:HIV-1 / Q000201:enzymology*; D007700:Kinetics; D008969:Molecular Sequence Data; D013379:Substrate Specificity
D015684:Gene Products, pol; D010450:Endopeptidases; D016333:HIV Protease