Proteases from human immunodeficiency virus and avian myeloblastosis virus show distinct specificities in hydrolysis of multidomain protein substrates.
The virally encoded proteases from human immunodeficiency virus (HIV) and avian myeloblastosis virus (AMV) have been compared relative to their ability to hydrolyze a variant of the three-domain Pseudomonas exotoxin, PE66. This exotoxin derivative, missing domain I and referred to as LysPE40, is made up of a 13-kilodalton NH2-terminal translocation domain II connected by a segment of 40 amino acids to enzyme domain III of the toxin, a 23-kilodalton ADP-ribosyltransferase. HIV protease hydrolyzes two peptide bonds in LysPE40, a Leu-Leu bond in the interdomain region and a Leu-Ala bond in a nonstructured region three residues in from the NH2-terminus. Neither of these sites is cleaved by the AMV enzyme; hydrolysis occurs, instead, at an Asp-Val bond in another part of the interdomain segment and at a Leu-Thr bond in the NH2-terminal region of domain II. Synthetic peptides corresponding to these cleavage sites are hydrolyzed by the individual proteases with the same specificity displayed toward the protein substrate. Peptide substrates for one protease are neither substrates nor competitive inhibitors for the other. A potent inhibitor of HIV type 1 protease was more than 3 orders of magnitude less active toward the AMV enzyme. These results suggest that although the crystallographic models of Rous sarcoma virus protease (an enzyme nearly identical to the AMV enzyme) and HIV type 1 protease show a high degree of similarity, there exist structural differences between these retroviral proteases that are clearly reflected by their kinetic properties.
Tomasselli
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AG
Biopolymer Chemistry Unit, Upjohn Company, Kalamazoo, Michigan 49008.
Hui
J O
JO
Sawyer
T K
TK
Staples
D J
DJ
Bannow
C A
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Reardon
I M
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Chaudhary
V K
VK
Fryling
C M
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Pastan
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I
Fitzgerald
D J
DJ
D036002:ADP Ribose Transferases; D000595:Amino Acid Sequence; D001354:Avian Leukosis Virus / Q000201:enzymology*; D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D001427:Bacterial Toxins; D015704:CD4 Antigens / Q000378:metabolism; D005098:Exotoxins / Q000378:metabolism; D015683:Gene Products, gag / Q000378:metabolism; D015684:Gene Products, pol / Q000378:metabolism; D015497:HIV-1 / Q000201:enzymology*; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism*; D010449:Peptide Mapping; D013379:Substrate Specificity; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D015704:CD4 Antigens; D005098:Exotoxins; D015683:Gene Products, gag; D015684:Gene Products, pol; D010447:Peptide Hydrolases; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A