Activity Staining and Inhibition Characterization of Dipeptidylpeptidase-III Enzyme from Goat Brain. (28th March 2011)

• Record Type: Journal Article
• Authors: Attri, Pooja; Singh, Jasbir; Dhanda, Suman; Singh, Hari
• Other Names: Nes William David Academic Editor.
• Is Part Of: Enzyme research. Volume 2011(2011)
• Publication Date: 2011-03-28
• Subjects: Enzymes -- Periodicals; 572.7
• DOI: 10.4061/2011/897028
• Languages: English
• ISSNs: 2090-0406

Abstract

Abstract : Dipeptidylpeptidase-III (DPP-III) from goat brain was purified and characterized using Arginyl-Arginyl-4-methoxy- β -naphthylamide (Arg-Arg-4m β NA) substrate. This enzyme retained its activity in native 10% polyacrylamide gel when stained using Arg-Arg-4m β NA. The activity was significantly increased by 100 mM chloride. Studies for its inhibition with some peptides and chemical inhibitors revealed that Leu-Trp-Met-Arg-Phe-Ala was most potent inhibitor followed by Arg-Phe-Ala and Gly-Phe-Leu. All the studied chemical inhibitors caused 40–50% inhibition at 1 mM. Metal ions helped to regain activity of EDTA pretreated enzyme. ZnCl2 at 50 μ M almost completely restored the enzyme activity. Further ZnCl2 and CoCl2 exerted protective effects on EDTA pretreated enzyme for its susceptibility to DTNB inhibition. Therefore, DPP-III is a metalloprotease with the involvement of cysteine residues either located at the catalytic site or involved in regulation.