Purification of an 80,000-Mr glycylprolyl peptidase from Bacteroides gingivalis.
An enzyme from Bacteroides gingivalis SUNYAB A7A1-28 that hydrolyzes the synthetic peptide glycyl-L-proline 4-methoxy-beta-naphthylamide was purified 1,040-fold by urea extraction, gel filtration, ion-exchange chromatography, and fast protein liquid chromatography. The molecular weight of the enzyme was 80,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 75,000 as determined by gel filtration. The optimum pH for the hydrolysis of glycyl-L-proline 4-methoxy-beta-naphthylamide was 7.5 to 8.5. The enzyme activity was inhibited by the serine protease inhibitors diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride by 82.5 and 78%, respectively. The activity was also inhibited by Hg2+ (55.6%) and Zn2+ (45%).
Barua
P K
PK
Department of Stomatology and Interdisciplinary Sciences, State University of New York, Buffalo 14214.
Neiders
M E
ME
Topolnycky
A
A
Zambon
J J
JJ
Birkedal-Hansen
H
H
D001426:Bacterial Proteins / Q000302:isolation & purification*; D001439:Bacteroides / Q000201:enzymology*; D003094:Collagen / Q000378:metabolism; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D008628:Mercury / Q000494:pharmacology; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D015032:Zinc / Q000494:pharmacology
D001426:Bacterial Proteins; D011480:Protease Inhibitors; D003094:Collagen; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C059607:glycylprolyl dipeptidase; D008628:Mercury; D015032:Zinc