Manganese-Induced Substrate Promiscuity in the Reaction Catalyzed by Phosphoglutamine Cytidylyltransferase from Campylobacter jejuni.

Abstract

The leading cause of bacterial gastroenteritis, Campylobacter jejuni, is a Gram-negative pathogen that contains a unique O-methyl phosphoramidate (MeOPN) on its capsular polysaccharide. Previously, MeOPN has been linked to the evasion of host immune responses and serum resistance. Despite the involvement of MeOPN in pathogenicity, the complete biosynthesis of this modification is unknown; however, the first four enzymatic steps have been elucidated. The second enzyme in this pathway, Cj1416, is a CTP/phosphoglutamine cytididylyltransferase that catalyzes the displacement of pyrophosphate from MgCTP by l-glutamine phosphate to form CDP-l-glutamine. Initially, Cj1416 was predicted to use phosphoramidate to form cytidine diphosphoramidate, but no activity was detected with MgATP as a substrate. However, in the presence of MnCTP, Cj1416 can directly catalyze the formation of cytidine diphosphoramidate from phosphoramidate and MnCTP. Here we characterize the manganese-induced promiscuity of Cj1416. In the presence of Mn2+, Cj1416 catalyzes the formation of 12 different reaction products using l-glutamine phosphate, phosphoramidate, methyl phosphate, methyl phosphonate, phosphate, arsenate, ethanolamine phosphate, glycerol-1-phosphate, glycerol-2-phosphate, serinol phosphate, l-serine phosphate, or 3-phospho-d-glycerate as the nucleophile to displace pyrophosphate from CTP.

• Authors: Zane W Taylor, Frank M Raushel
• Journal: Biochemistry (Biochemistry) Vol. 58 Issue 16 Pg. 2144-2151 (04 23 2019) ISSN: 1520-4995 [Electronic] United States
• PMID: 30929435 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Amides
• Bacterial Proteins
• Phosphoric Acids
• Glutamine
• Manganese
• Cytidine Triphosphate
• phosphoramidic acid
• Nucleotidyltransferases

Topics

• Amides (chemistry, metabolism)
• Bacterial Proteins (metabolism)
• Biocatalysis
• Campylobacter jejuni (enzymology)
• Cytidine Triphosphate (chemistry, metabolism)
• Glutamine (metabolism)
• Manganese (metabolism)
• Models, Chemical
• Molecular Structure
• Nucleotidyltransferases (metabolism)
• Phosphoric Acids (chemistry, metabolism)
• Substrate Specificity