TY - JOUR
T1 - Substrate specificity of a cationic peptidase from Bromelia hemisphaerica L.
AU - Cortés-Vázquez, Ma Isabel
AU - Muñoz-Sánchez, José Luis
AU - Briones-Martínez, Roberto
PY - 2008
Y1 - 2008
N2 - A cationic peptidase, named hemisphaericin-C, has been purified from the juice of Bromelia hemisphaerica fruits by ammonium sulfate precipitation, gel filtration on Sephadex G-75 and cationic exchange chromatography on carboxymethyl cellulose (CMC), to yield a single 24 kDa band on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which showed esterase and proteolytic activities. The esterase activity was inhibited by E-64, leupeptin, and cystatin, but not by EDTA. Characterization of the primary specificity of hemisphaericin-C showed activity towards substrates specific for chymotrypsin: N-succinyl-L-Phe-p-nitroanilide (PHEpNA) and N-benzoyl-L-Tyr-p-nitroanilide (TYRpNA), and those for trypsin: N-benzoyl-L-arg-p-nitroanilide (BApNA) to a lower degree. The higher selectivity, assessed by Vmax/Km, was obtained for PHEpNA, the substrate containing the aromatic lateral chain amino acid at the P1 position. The preference of hemisphaericin-C for PHEpNA gives a clue in the search for a chymotrypsin-like peptidase from a vegetal source.
AB - A cationic peptidase, named hemisphaericin-C, has been purified from the juice of Bromelia hemisphaerica fruits by ammonium sulfate precipitation, gel filtration on Sephadex G-75 and cationic exchange chromatography on carboxymethyl cellulose (CMC), to yield a single 24 kDa band on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which showed esterase and proteolytic activities. The esterase activity was inhibited by E-64, leupeptin, and cystatin, but not by EDTA. Characterization of the primary specificity of hemisphaericin-C showed activity towards substrates specific for chymotrypsin: N-succinyl-L-Phe-p-nitroanilide (PHEpNA) and N-benzoyl-L-Tyr-p-nitroanilide (TYRpNA), and those for trypsin: N-benzoyl-L-arg-p-nitroanilide (BApNA) to a lower degree. The higher selectivity, assessed by Vmax/Km, was obtained for PHEpNA, the substrate containing the aromatic lateral chain amino acid at the P1 position. The preference of hemisphaericin-C for PHEpNA gives a clue in the search for a chymotrypsin-like peptidase from a vegetal source.
KW - Bromelia hemisphaerica L.
KW - Cationic protease
KW - Hemisphaericin-C
KW - Trypsin and chymotrypsin-like specificity
UR - http://www.scopus.com/inward/record.url?scp=77953993134&partnerID=8YFLogxK
M3 - Artículo
SN - 1934-578X
VL - 3
SP - 351
EP - 355
JO - Natural Product Communications
JF - Natural Product Communications
IS - 3
ER -