Preparation and characterization of a truncated caricain lacking 41 residues from the N-terminal.
We purified an 18.8 kD protease from caricain solution. This protease was derived from caricain. It does not have the first 41 residues of the N-terminal sequence of caricain, and its N-terminal residue is Thr. Also, one of the disulfide bonds of caricain (cys22-cys63) was opened during the formation of the protease. We named this 18.8 kD protease caricain II. Caricain II has a wide pH range, and it is more sensitive to temperature changes than caricain. The proteolytic activity of caricain II is twice as much as that of caricain using casein as a substrate. However, caricain II has a low hydrolytic activity with N-benzoyl-L-arginine ethyl ester (BAEE) that is one of the special substrates of caricain. Our results indicate that caricain II is remarkably different from caricain and it can provide an improvement over caricain on the proteolytic activity.
Liu
Wei
W
South China Botanical of Garden, The Chinese Academy of Sciences, 510650, LeYiJu, Guangzhou, The People's Republic of China.
Ye
Wanhui
W
Wang
Zhangming
Z
Chao
Honglin
H
Lian
Juyu
J
D000595:Amino Acid Sequence; D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature
D002364:Caseins; D010446:Peptide Fragments; D010940:Plant Proteins; D001120:Arginine; C009727:benzoylarginine ethyl ester; D003546:Cysteine Endopeptidases; C507212:caricain II; C039974:caricain