SbnI is a free serine kinase that generates O -phospho-l-serine for staphyloferrin B biosynthesis in Staphylococcus aureus .

Autor:	Verstraete MM; From the Department of Microbiology and Immunology and., Perez-Borrajero C; the Genome Sciences and Technology Program Life Sciences Institute, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada and., Brown KL; From the Department of Microbiology and Immunology and., Heinrichs DE; the Department of Microbiology and Immunology, University of Western Ontario, London, Ontario N6A 5C1, Canada., Murphy MEP; From the Department of Microbiology and Immunology and michael.murphy@ubc.ca.
Jazyk:	angličtina
Zdroj:	The Journal of biological chemistry [J Biol Chem] 2018 Apr 20; Vol. 293 (16), pp. 6147-6160. Date of Electronic Publication: 2018 Feb 26.
DOI:	10.1074/jbc.RA118.001875
Abstrakt:	Staphyloferrin B (SB) is an iron-chelating siderophore produced by Staphylococcus aureus in invasive infections. Proteins for SB biosynthesis and export are encoded by the sbnABCDEFGHI gene cluster, in which SbnI, a member of the ParB/Srx superfamily, acts as a heme-dependent transcriptional regulator of the sbn locus. However, no structural or functional information about SbnI is available. Here, a crystal structure of SbnI revealed striking structural similarity to an ADP-dependent free serine kinase, SerK, from the archaea Thermococcus kodakarensis We found that features of the active sites are conserved, and biochemical assays and 31 P NMR and HPLC analyses indicated that SbnI is also a free serine kinase but uses ATP rather than ADP as phosphate donor to generate the SB precursor O -phospho-l-serine (OPS). SbnI consists of two domains, and elevated B -factors in domain II were consistent with the open-close reaction mechanism previously reported for SerK. Mutagenesis of Glu 20 and Asp 58 in SbnI disclosed that they are required for kinase activity. The only known OPS source in bacteria is through the phosphoserine aminotransferase activity of SerC within the serine biosynthesis pathway, and we demonstrate that an S. aureus serC mutant is a serine auxotroph, consistent with a function in l-serine biosynthesis. However, the serC mutant strain could produce SB when provided l-serine, suggesting that SbnI produces OPS for SB biosynthesis in vivo These findings indicate that besides transcriptionally regulating the sbn locus, SbnI also has an enzymatic role in the SB biosynthetic pathway. (© 2018 Verstraete et al.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu