Cloning, expression and genomic organization of human placental protein disulfide isomerase (previously identified as phospholipase C alpha).
Phosphoinositol-specific Phospholipase C plays an important role in transducing receptor generated signals to the rest of the cell. A cDNA encoding a phospholipase has been described (Bennett et al., 1988, Nature 334, 268-270). However it is probable that this cDNA in fact encodes a protein disulfide isomerase. Since the original work suggested that this enzyme was important in the reproductive tract we sort to clone, sequence, express and characterize the recombinant protein isolated from the placenta. We have cloned and sequenced the cDNA encoding the human homolog of this cDNA from human placenta, although the mRNA was widespread in the female reproductive tract. We have transiently expressed it in both COS cells and also 1BR fibroblasts. Cell lysates were assayed for increased phospholipase activity and protein disulfide activity. We describe the entire cDNA sequence which is highly conserved between species. We have also cloned a portion of the genomic gene and described the intron/exon boundaries. In vitro translation of this cDNA showed that it encoded a protein of 61 kD with a cleavable signal peptide. Transient expression showed the protein produced had no phospholipase activity but did show protein disulfide isomerase activity. The expression work shows that this cDNA indeed encodes a protein disulfide isomerase and not a phospholipase. The nucleotide sequence shows marked conservation of the coding and regulatory regions which may suggest that this enzyme has evolved to perform a highly specialized function.
Charnock-Jones
D S
DS
Department of Obstetrics and Gynaecology University, of Cambridge, Rosie Maternity Hospital, Cambridge, UK.
Day
K
K
Smith
S K
SK
D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D015894:Genome, Human; D006801:Humans; D007535:Isomerases / Q000235:genetics*; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D019704:Protein Disulfide-Isomerases
D018076:DNA, Complementary; D007535:Isomerases; D019704:Protein Disulfide-Isomerases