The Helicobacter pylori gene encoding phosphatidylserine synthase: Sequence, expression, and insertional mutagenesis

The Helicobacter pylori pss gene, coding for phosphatidylserine synthase (PSS), was cloned and sequenced in this study, A polypeptide of 237 amino acids was deduced from the PSS sequence, H. pylori PSS exhibits significant amino acid sequence identity with the PSS proteins found in the archaebacterium Methanococcus jannaschii, the gram-positive bacterium Bacillus subtilis, and the yeast Saccharomyces cerevisiae but none with its Escherichia coli counterpart, Expression of the putative pss gene in maxicells gale rise to a product of similar to 26 kDa, which is in agreement with the predicted molecular mass of 26.617 Da. A manganese-dependent PSS activity was found In the membrane fractions of the E. coli cells overexpressing the H. pylori pss gene product, This result indicates that this enzyme is a membrane-bound protein, a conclusion which is supported by the fact that the PSS protein contains several local hydrophobic segments which could form transmembrane helices. The pss gene was inactivated with a chloramphenicol acetyltransferase cassette on the plasmid. However, an isogenic pss gene-disrupted mutant of H. pylori UA802 could not be obtained, suggesting that this enzyme plays an essential role in the growth of this organism.