Abstract

PURIFICATION, CHARECTERIZATION & QUANTIFICATION OF TYPE II CLASS CGMP-DEPENDENT PROTEIN KINASE II (PKG-II) FROM RAT INTESTINAL MUCOSA

Kondal Reddy Y.*, Appaji D., K.V.M. krishna, Lakshmi SVVNSM

ABSTRACT

CGMP-dependent protein kinase (PKG-II) is a serine/threonine-specific protein kinase that is activated by cGMP. It phosphorylates a number of biologically important targets and is implicated in the regulation of smooth muscle relaxation, platelet function, sperm metabolism, cell division, and nucleic acid synthesis. A protein kinase type II was purified from rat intestinal mucosa using ammonium sulphate fractionation, ion exchange chromatography on DEAE and phosphocellulose and affinity chromatography on phosvitin- and casein-sepharose columns using AKTA-FPLC system. The Isolated PKG-II was characterized using western blot technique in comaprision with Native form of PKG-II. Enzymatic activity was performed successfully to correlate the functioning aspects of both Native and isolated PKG-II proteins.

Keywords: PKG-II, Phosphorylation, AKTA, rat intestinal mucosal cells, etc.