Biochem Biophys Res Commun. 1991 Mar 29;175(3):784-94. Unique Identifier
The 99 residue human immunodeficiency virus type 1 proteinase has been
expressed in Escherichia coli as part of an autocleaving fusion protein.
Expression of the fusion protein is toxic to the host cells, however
yields of the released proteinase have been improved by optimising
induction nad harvest times to increase culture biomass, and decrease
degradation of the proteinase. Soluble proteinase was extracted from
these cells by a simple and highly efficient three step process.
N-terminal sequence analysis confirms that the enzyme preparation is
highly pure and correctly autoprocessed. The proteinase cleaves peptide
substrate IGCTLNFPISPIETV between F and P at pH 6.0 with a Km of 310
microM and a Kcat of 14s-1. The enzyme is sensitive to its ionic
environment, showing stimulation of activity at high salt
concentrations, and shows a pH optimising 5.5.
Amino Acid Sequence Androgen-Binding Proteins Base Sequence
Chloramphenicol Acetyltransferase/*GENETICS Chromatography, Gel
Chromatography, High Pressure Liquid Chromatography, Ion Exchange
Cloning, Molecular Electrophoresis, Polyacrylamide Gel Escherichia
coli/GENETICS Gene Expression/DRUG EFFECTS HIV
Protease/*GENETICS/ISOLATION & PURIF/METABOLISM
HIV-1/ENZYMOLOGY/*GENETICS Isopropyl Thiogalactoside/PHARMACOLOGY
Kinetics Molecular Sequence Data Molecular Weight Oligonucleotide
Probes Plasmids Recombinant Fusion Proteins/ISOLATION & PURIF
Recombinant Proteins/ISOLATION & PURIF/METABOLISM Restriction Mapping