Inhibitory selectivity of canecystatin: a recombinant cysteine peptidase inhibitor from sugarcane.
The cDNA of a cystein peptidase inhibitor was isolated from sugarcane and expressed in Escherichia coli. The protein, named canecystatin, has previously been shown to exert antifungal activity on the filamentous fungus Trichoderma reesei. Herein, the inhibitory specificity of canecystatin was further characterized. It inhibits the cysteine peptidases from plant source papain (Ki =3.3nM) and baupain (Ki=2.1x10(-8)M), but no inhibitory effect was observed on ficin or bromelain. Canecystatin also inhibits lysosomal cysteine peptidases such as human cathepsin B (Ki=125nM), cathepsin K (Ki=0.76nM), cathepsin L (Ki=0.6nM), and cathepsin V (Ki=1.0nM), but not the aspartyl peptidase cathepsin D. The activity of serine peptidases such as trypsin, chymotrypsin, pancreatic, and neutrophil elastases, and human plasma kallikrein is not affected by the inhibitor, nor is the activity of the metallopeptidases angiotensin converting enzyme and neutral endopeptidase. This is the first report of inhibitory activity of a sugarcane cystatin on cysteine peptidases.
Oliva
Maria Luiza Vilela
ML
Department of Biochemistry, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua Três de Maio, 100, 04044-020 São Paulo, SP, Brazil. maysa.bioq@epm.br
Carmona
Adriana K
AK
Andrade
Sheila S
SS
Cotrin
Simone S
SS
Soares-Costa
Andrea
A
Henrique-Silva
Flavio
F
D015891:Cystatins / Q000737:chemistry*; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000145:classification*; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D010940:Plant Proteins; D011994:Recombinant Proteins; C466627:canecystatin protein, sugarcane; D003546:Cysteine Endopeptidases