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PMID: 2644644 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Activity of purified biosynthetic proteinase of human immunodeficiency virus on natural substrates and synthetic peptides.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 3 ·1989-02-00 ·Pages 807-11

Kräusslich HG, Ingraham RH, Skoog MT, Wimmer E, Pallai PV, Carter CA

Abstract

Retroviral capsid proteins and replication enzymes are synthesized as polyproteins that are proteolytically processed to the mature products by a virus-encoded proteinase. We have purified the proteinase of human immunodeficiency virus (HIV), expressed in Escherichia coli, to approximately 90% purity. The purified enzyme at a concentration of approximately 20 nM gave rapid, efficient, and specific cleavage of an in vitro synthesized gag precursor protein. Purified HIV proteinase also induced specific cleavage of five decapeptide substrates whose amino acid sequences corresponded to cleavage sites in the HIV polyprotein but not of a peptide corresponding to a cleavage site in another retrovirus. Competition experiments with different peptides allowed a ranking of cleavage sites. Inhibition studies indicated that the HIV proteinase was inhibited by pepstatin A with an IC50 of 0.7 microM.

MeSH Terms
Amino Acid Sequence Cloning, Molecular HIV/enzymology,genetics Kinetics Peptide Hydrolases/genetics,isolation & purification,metabolism Plasmids Protein Biosynthesis Recombinant Proteins/isolation & purification,metabolism Substrate Specificity Transcription, Genetic Viral Proteins
Chemicals
Recombinant Proteins Viral Proteins Peptide Hydrolases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kräusslich H G
Department of Microbiology, State University of New York, Stony Brook 11794.
Ingraham R H
Skoog M T
Wimmer E
Pallai P V
Carter C A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.3.807
Published
1989-02-00
Pages
807-11
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC286566
Subset
IM
Grants
NIAID NIH HHS · AI25993 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.3.807 PMC Full Text View journal details
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