Forward and reverse mutations affecting the kinetics and apparent molecular weight of mammalian HGPRT.

Chinese hamster cells selected for resistance to 8-azaguanine following mutagenesis have hypoxanthine-guanine phosphoribosyltransferase (HGPRT; E.C. 2.4.2.8) with characteristics compatible with different mutations in the structural gene for that enzyme. Using immunopurification and SDS-polyacrylamide electrophoresis, mutants producing antigenically active forms of the enzyme can be analyzed for changes in the molecular weight of HGPRT. Enzyme subunits from mutants RJK3 and RJK39 are reduced in molecular weight by an estimated 4 and 2%, respectively. HGPRT activity is not detectable in RJK39. The enzyme from RJK3 is active but has altered substrate binding properties. Enzymes from two other mutants with altered kinetic properties, RJK44 and RJK47, have normal molecular weights. The genetic alterations of RJK44 and 47 are probably missense mutations, while RJK3 and 39 might contain either deletions or mutations causing premature peptide chain termination. Somatic cell hybridization between RJK39 and a revertant of that strain with HGPRT of normal molecular weight revealed that the revertant probably arose by intragenic mutation rather than extragenic mutation or suppression.