ePrints@IIScePrints@IISc Home | About | Browse | Latest Additions | Advanced Search | Contact | Help

Purification and characterization of arginine decarboxylase from cucumber (Cucumis sativus) seedlings

Prasad, GL and Adiga, PR (1985) Purification and characterization of arginine decarboxylase from cucumber (Cucumis sativus) seedlings. In: Journal of Biosciences, 7 (3-4). pp. 331-343.

[img] PDF
Purification_and_characterization.pdf - Published Version
Restricted to Registered users only

Download (719kB) | Request a copy
Official URL: http://www.springerlink.com/content/a075l426366322...

Abstract

A simple, reproducible and rapid protocol for the purification of arginine decarboxylase fromCucumis sativus seedlings has been standardised. The purification steps involved ion-exchange chromatography on diethylaminoethyl-cellulose followed by gel filtration on Sephadex G-l 50. The purified enzyme preparation migrated as a single stainable band on Polyacrylamide gels at both basic and acidic pH, but under denaturing and reducing conditions on sodium dodecyl sulphate-polyacrylamide gels resolved into polypeptides of molecular weight 48,000,44,000 and 15,000. However, in the absence of 2-mercaptoethanol on electrophoresis on sodium dodecyl sulphate-polyacrylamide gels, the enzyme moved as single band with a molecular weight of 150,000. Evidence was obtained to indicate that these three polypeptides were probably derived from a single larger molecular weight enzyme. On storage of the purified protein, the 48,000 species was preferentially degraded to smaller polypeptides. The preliminary data suggested that the 48,000 and 44,000 species shared many common tryptic peptides as revealed by finger printing of the $^{125}I$-labelled protein. The purified enzyme was a glycoprotein and had $aK_m$ of 0.5 mM for arginine. Its activity was stimulated by dithiothrietol and pyridoxal phosphate. EDTA did not inhibit the enzyme activity. $Mn^{2+}$ at 1 mM stimulated arginine decarboxylase activity but was inhibitory at higher concentration.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to Springer.
Keywords: Arginine decarboxylase;peptide mapping;purification
Department/Centre: Division of Biological Sciences > Biochemistry
Depositing User: RM Shivakumara
Date Deposited: 25 Jun 2008
Last Modified: 01 Mar 2012 06:10
URI: http://eprints.iisc.ac.in/id/eprint/14683

Actions (login required)

View Item View Item