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Volume 61 Number 4 >

Expression and characterization of cathepsin B from tsetse (Glossina morsitans morsitans)

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/jjvr.61.4.137

Title: Expression and characterization of cathepsin B from tsetse (Glossina morsitans morsitans)
Authors: Ruttayaporn, Ngasaman Browse this author
Zhou, Mo Browse this author
Suganuma, Keisuke Browse this author
Yamasaki, Shino Browse this author
Kawazu, Shin-ichiro Browse this author
Inoue, Noboru Browse this author
Keywords: cathepsin B
tsetse fly
enzyme assay
Issue Date: Nov-2013
Publisher: Graduate School of Veterinary Medicine, Hokkaido University
Journal Title: Japanese Journal of Veterinary Research
Volume: 61
Issue: 4
Start Page: 137
End Page: 147
Abstract: Digestive enzymes in tsetse fly midgut are thought to modulate the development of African trypanosome which is a causative agent of trypanosomosis in human and animal. Cathepsin B is induced after the first blood meal ingestion and being higher in trypanosome infected flies. A DNA fragment encoding pro-cathepsin B (930 bp) (Accession No. AF329480_1) was cloned and expressed in E. coli and P. pastoris protein expression systems. An active recombinant cathepsin B (rGmcathB) produced by P. pastoris was migrating from 35 to 45 kDa under reducing condition. rGmcathB exhibited the highest proteolytic activity at pH 4.0 with wide range temperature 25-30oC, also degraded bovine hemoglobin and serum albumin. rGmcathB exhibited hydrolysis preference for Z-Arg-Arg-MCA (Kcat/KM 7.58 mM-1sec-1) and bovine hemoglobin (Kcat/KM 3.77x103 mM-1sec-1). The proteolytic activity of rGmcathB was inhibited by specific cysteine protease inhibitor (E-64) confirmed belonging to papain-like cysteine protease family. These results indicated that rGmcathB shows the activity of cathepsin B and have high affinity with blood protein referring a role in blood meal digestion. In this study, the recombinant protein expressed by E. coli expression system was not enzymatically active as shown in the recombinant protein expressed by P. pastoris expression system. This finding implies that P. pastoris expression system is more suitable for expressing enzymatically active recombinant proteases than E. coli expression system.
Type: bulletin (article)
URI: http://hdl.handle.net/2115/53708
Appears in Collections:Japanese Journal of Veterinary Research > Volume 61 Number 4

Submitter: 獣医学部図書室

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