Crystal Structure of a Hidden Protein, YcaC, a Putative Cysteine Hydrolase from Pseudomonas aeruginosa, with and without an Acrylamide Adduct

Grøftehauge, M.K.; Truan, D.; Vasil, A.; Denny, P.W.; Vasil, M.L.; Pohl, E.

doi:10.3390/ijms160715971

Crystal Structure of a Hidden Protein, YcaC, a Putative Cysteine Hydrolase from Pseudomonas aeruginosa, with and without an Acrylamide Adduct

Grøftehauge, M.K.; Truan, D.; Vasil, A.; Denny, P.W.; Vasil, M.L.; Pohl, E.

Authors

M.K. Grøftehauge

D. Truan

A. Vasil

Professor Paul Denny p.w.denny@durham.ac.uk
Professor

M.L. Vasil

Professor Ehmke Pohl ehmke.pohl@durham.ac.uk
Professor

Abstract

As part of the ongoing effort to functionally and structurally characterize virulence factors in the opportunistic pathogen Pseudomonas aeruginosa, we determined the crystal structure of YcaC co-purified with the target protein at resolutions of 2.34 and 2.56 Å without a priori knowledge of the protein identity or experimental phases. The three-dimensional structure of YcaC adopts a well-known cysteine hydrolase fold with the putative active site residues conserved. The active site cysteine is covalently bound to propionamide in one crystal form, whereas the second form contains an S-mercaptocysteine. The precise biological function of YcaC is unknown; however, related prokaryotic proteins have functions in antibacterial resistance, siderophore production and NADH biosynthesis. Here, we show that YcaC is exceptionally well conserved across both bacterial and fungal species despite being non-ubiquitous. This suggests that whilst YcaC may not be part of an integral pathway, the function could confer a significant evolutionary advantage to microbial life.

Citation

Grøftehauge, M., Truan, D., Vasil, A., Denny, P., Vasil, M., & Pohl, E. (2015). Crystal Structure of a Hidden Protein, YcaC, a Putative Cysteine Hydrolase from Pseudomonas aeruginosa, with and without an Acrylamide Adduct. International Journal of Molecular Sciences, 16(7), 15971-15984. https://doi.org/10.3390/ijms160715971

Journal Article Type	Article
Acceptance Date	Jun 15, 2015
Publication Date	Jul 14, 2015
Deposit Date	Aug 25, 2015
Publicly Available Date	Sep 4, 2015
Journal	International Journal of Molecular Sciences
Print ISSN	1661-6596
Publisher	MDPI
Peer Reviewed	Peer Reviewed
Volume	16
Issue	7
Pages	15971-15984
DOI	https://doi.org/10.3390/ijms160715971
Keywords	X-ray crystallography, Micro-crystals, Molecular replacement, YcaC, Isochorismate family, Protein octamer.

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