Molecular model of equine infectious anemia virus proteinase and kinetic measurements for peptide substrates with single amino acid substitutions.
A molecular model has been built of the equine infectious anemia virus (EIAV) proteinase on the basis of the crystal structures of the related Rous sarcoma virus (RSV) and human immunodeficiency virus (HIV) proteinases. The 104 residue long EIAV proteinase has 30 identical and 11 similar amino acids compared to those in HIV-1 proteinase and 25 identical and 18 similar amino acids compared to RSV proteinase. The overall structure is predicted to be close to that of HIV-1 proteinase. Two regions show differences: there are 6 additional residues leading to the tip of the flap, which is predicted to be involved in interactions with substrate, and there is a single residue deletion in the beta b' strand at a position equivalent to residue 60 in HIV-1 proteinase. The conformation of the residues leading to the flap was modeled by analogy to the corresponding region of RSV proteinase. The peptide substrate, VSQNYPIVQ, was modeled by analogy to the inhibitors in the co-crystal structures of HIV-1 proteinase, and the residues forming the substrate binding sites of EIAV proteinase were identified. EIAV proteinase showed several non-conservative substitutions in these residues compared to HIV-1 proteinase: Thr 30 instead of Asp in subsites S2, S2', S4, and S4', Ile 54 instead of Gly 48 in subsites S1, S1', S3, and S3', Arg 79 instead of Thr 74 in S4 and S4', and Ile 85 instead of Thr 80 in subsites S1 and S1'.(ABSTRACT TRUNCATED AT 250 WORDS)
Weber
I T
IT
Department of Pharmacology, Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Tözsér
J
J
Wu
J
J
Friedman
D
D
Oroszlan
S
S
D000595:Amino Acid Sequence; D001665:Binding Sites; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D016333:HIV Protease / Q000737:chemistry; D004860:Infectious Anemia Virus, Equine / Q000201:enzymology*; D007700:Kinetics; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D013379:Substrate Specificity
D010455:Peptides; D010450:Endopeptidases; D016333:HIV Protease