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2016 Volume 7 Issue 1

Purification and characterization of cathepsin B from buffalo (Bubalus bubalis) lung


Abstract
In an attempt to improve the level of purity of buffalo lung cathepsin B, we have modified the earlier procedure by incorporating CM- Sephadex C-50 ion exchange chromatography and rechromatography on Sephacryl S-200 column. Appearance of a single band on SDS-PAGE under reducing and non-reducing conditions showed that the enzyme exists in a single chain molecule. Molecular weight of purified cathepsin B was determined to be 23800 and 25400 by SDS-PAGE and gel filtration, respectively. The enzyme, a glycoprotein, showed activity against α-N-benzoyl-DL-arginine-2-naphthylamide and α-N-benzoyl-DL-arginine-4-nitroanilide. The physico-chemical properties of the enzyme were similar to the properties reported for cathepsin B from other sources. However, the NH2-terminal amino acid residue of the enzyme was found to be Ala as against Leu reported from other sources. The enzyme was activated by various thiol reducing reagents and inhibited by cysteine protease inhibitors and denaturing agents. The hydrodynamic behaviour of cathepsin B which includes Stokes radius (2.29 nm), frictional ratio (1.19) and intrinsic viscosity (3.08 ml/gm) suggested that the native enzyme conformation is compact and globular. 

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JOURNAL OF BIOCHEMICAL TECHNOLOGY
JOURNAL OF BIOCHEMICAL TECHNOLOGY
Journal of Biochemical Technology is a peer reviewed International Journal published by the Sevas Publishing on behalf of the Biochemical Technology Society, a Registered Charity Organization from India

AREA OF INTEREST
AREA OF INTEREST
Areas of high interest, cover new advances in enzymatic and protein mechanims; applied molecular genetics and biotechnology; genomics and proteomics; metabolic; medical, environmental, food and agro biotechnology.

FOCUS AND SCOPE
FOCUS AND SCOPE
Journal Of Biochemical Technology” Provides A Medium For The Rapid Publication Of Full-Length Articles, Mini-Reviews Of New And Emerging Products And Short Communications On All Aspects Of ...

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This journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. Keywords include, Biochemical Research: Endo/exocytosis, Trafficking, Membrane Biology, Cell Migration, Cell-Matrix Organelle Biogenesis, Cytoskeleton Proteolysis, Cell Death, Cell Cycle, Cancer, Cell Growth/Death, Differentiation, Drug Targets, Gene Therapy, Models of Disease, Proteomics, Stem Cells, Bioenergetics, Mitochondria, Free Radicals, Redox Signaling, Ion Transport/Channels, Oxidative