A soluble active mutant of HIV-1 integrase: involvement of both the core and carboxyl-terminal domains in multimerization.

Important note: Information in this article was accurate in 1996. The state of the art may have changed since the publication date.

A soluble active mutant of HIV-1 integrase: involvement of both the core and carboxyl-terminal domains in multimerization.

J Biol Chem. 1996 Mar 29;271(13):7712-8. Unique Identifier : AIDSLINE MED/96205962
Jenkins TM; Engelman A; Ghirlando R; Craigie R; Laboratory of Molecular Biology, NIDDK, National Institutes of; Health, Bethesda, Maryland 20892-0560, USA.

Abstract: Structural studies of human immunodeficiency virus type 1 (HIV-1) integrase have been impeded by the low solubility of the protein. By systematic replacement of hydrophobic residues, we previously identified a single amino acid change (F185K) that dramatically improved the solubility of the catalytic domain of HIV-1 integrase and enabled the structure to be determined by x-ray crystallography. We have introduced the same mutation into full-length HIV-1 integrase. The resulting recombinant protein is soluble and fully active in vitro, whereas, HIV-1 carrying the mutation is replication-defective due to improper virus assembly. Analysis of the recombinant protein by gel filtration and sedimentation equilibrium demonstrate a dimer-tetramer self-association. We find that the regions involved in multimerization map to both the catalytic core and carboxyl-terminal domains. The dramatically improved solubility of this protein make it a good candidate for structural studies.

Keywords: Amino Acid Sequence Base Sequence Comparative Study DNA Nucleotidyltransferases/BIOSYNTHESIS/*CHEMISTRY/*METABOLISM Electrophoresis, Polyacrylamide Gel Human HIV-1/*ENZYMOLOGY/GENETICS/PHYSIOLOGY Kinetics Macromolecular Systems Molecular Sequence Data Mutagenesis, Site-Directed Oligodeoxyribonucleotides Point Mutation Recombinant Proteins/BIOSYNTHESIS/CHEMISTRY/METABOLISM Solubility Substrate Specificity Ultracentrifugation Virus Integration Virus Replication JOURNAL ARTICLE
960930
M9690913

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