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Proc Natl Acad Sci U S A. 2015 Apr 21;112(16):E1974-83. doi: 10.1073/pnas.1423570112. Epub 2015 Apr 6.

Panoramic view of a superfamily of phosphatases through substrate profiling.

Author information

1
Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
2
Department of Chemistry, Boston University, Boston, MA 02215; Bioinformatics Graduate Program, Boston University, Boston, MA 02215; and.
3
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
4
Department of Chemistry, Boston University, Boston, MA 02215;
5
Department of Chemistry, Boston University, Boston, MA 02215; drkallen@bu.edu jfarelli@bu.edu.

Abstract

Large-scale activity profiling of enzyme superfamilies provides information about cellular functions as well as the intrinsic binding capabilities of conserved folds. Herein, the functional space of the ubiquitous haloalkanoate dehalogenase superfamily (HADSF) was revealed by screening a customized substrate library against >200 enzymes from representative prokaryotic species, enabling inferred annotation of ∼35% of the HADSF. An extremely high level of substrate ambiguity was revealed, with the majority of HADSF enzymes using more than five substrates. Substrate profiling allowed assignment of function to previously unannotated enzymes with known structure, uncovered potential new pathways, and identified iso-functional orthologs from evolutionarily distant taxonomic groups. Intriguingly, the HADSF subfamily having the least structural elaboration of the Rossmann fold catalytic domain was the most specific, consistent with the concept that domain insertions drive the evolution of new functions and that the broad specificity observed in HADSF may be a relic of this process.

KEYWORDS:

evolution; phosphatase; promiscuity; specificity; substrate screen

PMID:
25848029
PMCID:
PMC4413258
DOI:
10.1073/pnas.1423570112
[Indexed for MEDLINE]
Free PMC Article

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