Analysis of a Bacillus subtilis proteinase mutant.
A Bacillus subtilis mutant having a phenotype manifesting reduced extracellular proteolytic activity was investigated. An extracellular protein was isolated and shown by fingerprint analysis to be a fragment of the wild-type enzyme. By using previously established molecular weights for the wild-type enzyme (2.9 x 10(4)) and the two polypeptide chains derived from it (1.4 x 10(4) each), with the amino acid analysis and fingerprints of both wild-type and mutant proteins, a molecular weight of 1.57 x 10(4) was assigned to the mutant protein. (32)P-diisopropylphosphate labeling of the mutant protein showed only 1 in 53 molecules to be functional. Thin-layer chromatography on Sephadex G-75 demonstrated that the active molecules were separable from the bulk of the isolated protein and had the same mobility as the wild-type enzyme. Fingerprints of tryptic digests of (32)P-diisopropylphosphate-labeled wild-type and mutant proteins showed that the labeled peptides had identical characteristics.
D000596:Amino Acids / Q000032:analysis; D001322:Autoanalysis; D001412:Bacillus subtilis / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism*; D002247:Carbon Isotopes; D002474:Cell-Free System; D002855:Chromatography, Thin Layer; D004589:Electrophoresis, Disc; D005827:Genetics, Microbial; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009154:Mutation; D010447:Peptide Hydrolases / Q000032:analysis / Q000378:metabolism*; D010455:Peptides / Q000032:analysis; D010641:Phenotype; D010710:Phosphates / Q000378:metabolism; D010759:Phosphorus Isotopes; D014357:Trypsin; D014461:Ultracentrifugation
D000596:Amino Acids; D001426:Bacterial Proteins; D002247:Carbon Isotopes; D010455:Peptides; D010710:Phosphates; D010759:Phosphorus Isotopes; D010447:Peptide Hydrolases; D014357:Trypsin