Affinity purification of HIV-1 and HIV-2 proteases from recombinant E. coli strains using pepstatin-agarose
Biochemical and Biophysical Research CommunicationsPublished 1 August 1990
Judith Rittenhouse, Mary C. Turon, Rosalind Helfrich, Kathryn S. Albrecht, Debra Weigl, Robert L. Simmer
Citations34
SJR quartileQ2
SJR score0.75
SNIP0.56
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TL;DR
A procedure is described which employs pepstatin-agarose for the affinity purification of either HIV-1 or HIV-2 protease from two similar recombinant E. coli constructs that were developed for the expression of these enzymes.
Abstract
A procedure is described which employs pepstatin-agarose for the affinity purification of either HIV-1 or HIV-2 protease from two similar recombinant E. coli constructs that were developed for the expression of these enzymes. HIV-2 protease was routinely expressed at much higher levels than the HIV-1 enzyme and pepstatin-agarose was the only chromatography step required to isolate pure HIV-2 protease from crude bacterial lysates. A Mono S ionic exchange step following pepstatin-agarose chromatography was sufficient to bring the HIV-1 protease to homogeneity. Purification of either enzyme can be completed in several days yielding homogeneous preparations suitable for crystallization and other physical characterization.
Keywords
Immunology and MicrobiologyMedicine
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