Bacillopeptidase F: two forms of a glycoprotein serine protease from Bacillus subtilis 168.
Bacillopeptidase F is a serine endopeptidase excreted by Bacillus subtilis 168 after the end of exponential growth. As a step toward discovering a physiological function for this protease, an enzymological and immunological study was undertaken. When bacillopeptidase F was purified at pH 10, a number of enzymically active, rapidly moving electrophoretic forms were observed, as had been previously reported. Rabbit antiserum was prepared against one form. When the enzyme was purified at pH 6.0 in the presence of the covalent inhibitor phenylmethylsulfonyl fluoride, using the rabbit antiserum to detect the bacillopeptidase F protein, no fast-moving electrophoretic forms were observed. Instead, only two forms of the enzyme were isolated. One form had a molecular weight of 33,000, and the other had a molecular weight of 50,000, as determined by equilibrium sedimentation methods. Both forms appeared to be glycoproteins, both contained compounds, released on acid hydrolysis, which cochromatographed with phosphoserine and galactosamine, and the two gave identical immunoprecipitin lines in Ouchterlony double-diffusion tests. The smaller form had a pI of 4.4, whereas the larger had a pI of 5.4. The data suggest that bacillopeptidase F is distinct from all other proteases of B. subtilis.
D000596:Amino Acids / Q000032:analysis; D001412:Bacillus subtilis / Q000201:enzymology* / Q000254:growth & development; D002241:Carbohydrates / Q000032:analysis; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D006023:Glycoproteins / Q000302:isolation & purification; D005779:Immunodiffusion; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D012697:Serine Endopeptidases
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D007527:Isoenzymes; D010450:Endopeptidases; D012697:Serine Endopeptidases; C038477:bacillopeptidase F