Cloning and Expression of Pleckstrin 2, a Novel Member of the Pleckstrin Family

Abstract

A novel member of the pleckstrin family has been identified and designated as mouse pleckstrin 2. The cDNA clone with an insert of 1588 bp contained a 1059-bp open reading frame encoding a polypeptide of 353 amino acid residues. The deduced amino acid sequence predicted that pleckstrin 2 has two pleckstrin homology (PH) domains at the N- and C-termini and a DEP (Dsh, egl-10, and pleckstrin) domain in the central region and showed 35% identity with the sequence of chicken pleckstrin. Northern blot and reverse-transcription polymerase chain reaction analysis revealed that pleckstrin 2 mRNA is ubiquitously expressed. Southern blot analysis indicated that the mouse pleckstrin 2 gene may consisit of two or more exons. To obtain information relative to natural ligand(s) for each of the PH domains in vivo, we employed the green fluorescent protein (GFP) tagged fusion protein system. Distributions of N-terminal and C-terminal PH domains of pleckstrin 2 were quite different from each other, suggesting that these PH domains may interact with distinct factor(s).