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Evaluation of synthetic thiosemicarbazone containing compounds as inhibitors of human recombinant cathepsin B
Author(s) -
Lucas Joseph Anthony,
Yelma Niranjan Reddy,
Wen Lisa,
Huang Jenq-Kuen,
McConnell Rose
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.956.4
Subject(s) - cathepsin b , recombinant dna , chemistry , cathepsin , biochemistry , cysteine protease , enzyme , inclusion bodies , microbiology and biotechnology , protease , biology , gene
The objective of this study was to test the inhibitory effects of eight different synthetic compounds on recombinant Cathepsin B, a lysosomal cysteine protease that plays roles in processes such as protein degradation, bone resorption, and antigen processing. This protein has been found in abnormally high levels in the bloodstream of both Alzheimer's and cancer patients. In order to investigate the veracity of synthetic Cathepsin B inhibitor candidates, recombinant procathepsin B protein was obtained by overexpression of previously constructed E. coli transformant followed by purification of inclusion bodies, solubilization, and refolding. After refolding, the procathepsin B was autoactivated by lowing the pH to 4 with formic acid. Finally, the effectiveness of each inhibitor candidate was determined via colorimetric enzymatic assay using N‐Carbobenzoxy‐L‐Phenylalanyl‐Arginine‐4‐nitroanilide hydrochloride as the chromogenic substrate. The results of each inhibitor candidate are presented as an IC 50 (50% Inhibitory Concentration). Funding provided by National Cancer Institute at NIH (Grant No. 3R15CA086933‐04 and 3R15CA086933‐04A2S1) and Western Illinois University.