title: Activity Staining and Inhibition Characterization of Dipeptidylpeptidase-III Enzyme from Goat Brain.
abstract: Dipeptidylpeptidase-III (DPP-III) from goat brain was purified and characterized using Arginyl-Arginyl-4-methoxy-β-naphthylamide (Arg-Arg-4mβNA) substrate. This enzyme retained its activity in native 10% polyacrylamide gel when stained using Arg-Arg-4mβNA. 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. ZnCl(2) at 50 μM almost completely restored the enzyme activity. Further ZnCl(2) and CoCl(2) 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.
authors: Department of Biochemistry, Kurukshetra University, Haryana, Kurukshetra 136119, India.
authors: Pooja
authors: P
authors: Attri
authors: Jasbir
authors: J
authors: Singh
authors: Suman
authors: S
authors: Dhanda
authors: Hari
authors: H
authors: Singh
references: Protein Expr Purif. 2007 Apr;52(2):297-305
references: 17182254
references: Chem Pharm Bull (Tokyo). 1986 Aug;34(8):3333-40
references: 3791505
references: J Neurochem. 1994 Oct;63(4):1439-45
references: 7931295
references: Ann N Y Acad Sci. 1964 Dec 28;121:404-27
references: 14240539
references: J Biochem. 1984 Jul;96(1):107-15
references: 6490598
references: Masui. 2003 Mar;52(3):257-63
references: 12703067
references: J Biochem. 1983 Jul;94(1):267-73
references: 6619113
references: Cell Biochem Funct. 2008 Apr;26(3):339-45
references: 18064728
references: J Enzyme Inhib Med Chem. 2008 Apr;23(2):174-81
references: 18343901
references: Biol Chem Hoppe Seyler. 1988 Jan;369(1):29-38
references: 3348886
references: J Biol Chem. 1967 Oct 25;242(20):4623-9
references: 6061409