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Lookup NU author(s): Dr Christopher Johnson, Dr Heather Lamb, Professor Alastair Hawkins
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The Salmonella typhimurium "yeaZ" gene (StyeaZ) encodes an essential protein of unknown function (StYeaZ), which has previously been annotated as a putative homolog of the Pasteurella haemolytica M22 O-sialoglycoprotein endopeptidase Gcp. YeaZ has also recently been reported as the first example of an RPF from a gram-negative bacterial species. To further characterize the properties of StYeaZ and the widely occurring MK-M22 family, we describe the purification, biochemical analysis, crystallization, and structure determination of StYeaZ. The crystal structure of StYeaZ reveals a classic two-lobed actin-like fold with structural features consistent with nucleotide binding. However, microcalorimetry experiments indicated that StYeaZ neither binds polyphosphates nor a wide range of nucleotides. Additionally, biochemical assays show that YeaZ is not an active O-sialoglycoprotein endopeptidase, consistent with the lack of the critical zinc binding motif. We present a detailed comparison of YeaZ with available structural homologs, the first reported structural analysis of an MK-M22 family member. The analysis indicates that StYeaZ has an unusual orientation of the A and B lobes which may require substantial relative movement or interaction with a partner protein in order to bind ligands. Comparison of the fold of YeaZ with that of a known RPF domain from a gram-positive species shows significant structural differences and therefore potentially distinctive RPF mechanisms for these two bacterial classes. © 2006 Wiley-Liss, Inc.
Author(s): Nichols CE, Johnson C, Lockyer M, Charles IG, Lamb HK, Hawkins AR, Stammers DK
Publication type: Article
Publication status: Published
Journal: Proteins: Structure, Function and Genetics
Year: 2006
Volume: 64
Issue: 1
Pages: 111-123
ISSN (print): 0887-3585
ISSN (electronic): 1097-0134
Publisher: John Wiley & Sons, Inc.
URL: http://dx.doi.org/10.1002/prot.20982
DOI: 10.1002/prot.20982
PubMed id: 16617437
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