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PLoS One. 2014 Sep 19;9(9):e108310. doi: 10.1371/journal.pone.0108310. eCollection 2014.

Common hydrogen bond interactions in diverse phosphoryl transfer active sites.

Author information

1
Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health and Science University, Portland, Oregon, United States of America.
2
Center for Computational Sciences and the Department of Chemistry and Biochemistry, Duquesne University, Pittsburgh, Pennsylvania, United States of America.

Abstract

Phosphoryl transfer reactions figure prominently in energy metabolism, signaling, transport and motility. Prior detailed studies of selected systems have highlighted mechanistic features that distinguish different phosphoryl transfer enzymes. Here, a top-down approach is developed for comparing statistically the active site configurations between populations of diverse structures in the Protein Data Bank, and it reveals patterns of hydrogen bonding that transcend enzyme families. Through analysis of large samples of structures, insights are drawn at a level of detail exceeding the experimental precision of an individual structure. In phosphagen kinases, for example, hydrogen bonds with the O3β of the nucleotide substrate are revealed as analogous to those in unrelated G proteins. In G proteins and other enzymes, interactions with O3β have been understood in terms of electrostatic favoring of the transition state. Ground state quantum mechanical calculations on model compounds show that the active site interactions highlighted in our database analysis can affect substrate phosphate charge and bond length, in ways that are consistent with prior experimental observations, by modulating hyperconjugative orbital interactions that weaken the scissile bond. Testing experimentally the inference about the importance of O3β interactions in phosphagen kinases, mutation of arginine kinase Arg280 decreases kcat, as predicted, with little impact upon KM.

PMID:
25238155
PMCID:
PMC4169622
DOI:
10.1371/journal.pone.0108310
[Indexed for MEDLINE]
Free PMC Article

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