title: beta-Galactosidase containing a human immunodeficiency virus protease cleavage site is cleaved and inactivated by human immunodeficiency virus protease.
abstract: A "cleavage cassette" specifying a decapeptide human immunodeficiency virus (HIV) protease cleavage site was introduced into six different locations of beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) in Escherichia coli. Four of these constructs retained beta-galactosidase activity despite the insertion of the cleavage cassette. Of these four constructs, one was cleaved by HIV protease, resulting in the inactivation of beta-galactosidase both in vivo and in vitro. This cleavage was inhibited by pepstatin A, a known inhibitor of HIV protease. Thus, beta-galactosidase has been converted into an easily assayed substrate for HIV protease. An analogous construct of beta-galactosidase containing a polio protease cleavage site was cleaved likewise by polio protease, suggesting that this system may be generic for monitoring cleavage by a variety of proteases.
authors: Molecular Biology Section, Lederle Laboratories, American Cyanamid Company, Pearl River, NY 10965.
authors: E Z
authors: EZ
authors: Baum
authors: G A
authors: GA
authors: Bebernitz
authors: Y
authors: Y
authors: Gluzman
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D016333:HIV Protease / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010436:Pepstatins / Q000494:pharmacology; D010957:Plasmids; D015183:Restriction Mapping; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D001616:beta-Galactosidase; D016333:HIV Protease; C031375:pepstatin