Expression, purification and characterisation of a functional phosphatidylinositol‐specific phospholipase C‐δ1 protein in Escherichia coli

Abstract

Inositol‐lipid‐specific phospholipase C‐δ1 (PtdIns‐PLCδ1) was expressed in Escherichia coli as a fusion protein containing a short 22‐amino‐acid lac‐Z‐derived amino terminus. Under appropriate conditions, the phospholipase constituted approximately 0.2% of the detergent‐soluble protein and could be purified to near homogeneity in a simple three step protocol. The catalytic properties of the purified enzyme closely resemble those of the eukaryote‐derived protein. The suitability of bacterial expression for the investigation of PtdIns‐PLCδ regulation is discussed. Copyright © 1992, Wiley Blackwell. All rights reserved