Kinetic characterization of gyroxin, a serine protease from Crotalus durissus terrificus venom.
This work describes for the first time the characterization of the enzymatic features of gyroxin, a serine protease from Crotalus durissus terrificus venom, capable to induce barrel rotation syndrome in rodents. Measuring the hydrolysis of the substrate ZFR-MCA, the optimal pH for proteolytic cleavage of gyroxin was found to be at pH 8.4. Increases in the hydrolytic activity were observed at temperatures from 25 °C to 45 °C, and increases of NaCl concentration up to 1 M led to activity decreases. The preference of gyroxin for Arg residues at the substrate P1 position was also demonstrated. Taken together, this work describes the characterization of substrate specificity of gyroxin, as well as the effects of salt and pH on its enzymatic activity.
Depto Farmacologia, UNIFESP, 04044-020 São Paulo-SP, Brazil.
Camila M
CM
Yonamine
Marcia Y
MY
Kondo
Maria A
MA
Juliano
Marcelo Y
MY
Icimoto
Gandhi R
GR
Baptista
Tetsuo
T
Yamane
Vitor
V
Oliveira
Luis
L
Juliano
Antônio J
AJ
Lapa
Maria Teresa R
MT
Lima-Landman
Mirian A F
MA
Hayashi
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D055162:Biocatalysis / Q000187:drug effects; D002942:Circular Dichroism; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D017839:Crotalus / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D057057:Serine Proteases / Q000737:chemistry / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013696:Temperature
D003435:Crotalid Venoms; D009498:Neurotoxins; D010455:Peptides; C017356:gyroxin; D012965:Sodium Chloride; D001120:Arginine; D057057:Serine Proteases