Affinity purification of the HIV-1 protease

Biochemical and Biophysical Research CommunicationsPublished 1 November 1989

Jill C. Heimbach, Victor M. Garsky, Stuart R. Michelson, Richard A. F. Dixon, Irving S. Sigal, Paul L. Darke

DOI10.1016/0006-291x(89)91762-2

Source URL

Citations58

SJR quartileQ2

SJR score0.75

SNIP0.56

TL;DR

An inhibitor of the HIV-1 protease has been employed in the generation of a resin which allows the rapid purification of this enzyme in a homogeneous preparation capable of crystallization.

Abstract

An inhibitor of the HIV-1 protease has been employed in the generation of a resin which allows the rapid purification of this enzyme. A peptide substrate analogue, H2N-Ser-Gln-Asn-(Phe-psi[CH2N]-Pro)-Ile-Val-Gln-OH, was coupled to agarose resin. The HIV-1 protease was expressed in E. coli and the supernatant from lysed cells was passed through the affinity resin. Active HIV-1 protease was then eluted with a buffer change to pH 10 and 2 M NaCl. Final purification to a homogeneous preparation, capable of crystallization, was achieved with hydrophobic interaction chromatography. Solutions containing HIV-1 protease bound to competitive inhibitors do not bind to the column.

Keywords

Immunology and MicrobiologyMedicine

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