Panoramic view of a superfamily of phosphatases through substrate profiling

Hua Huang, Chetanya Pandya, Chunliang Liu, Nawar F. Al-Obaidi, Min Wang, Li Zheng, Sarah Toews Keating, Miyuki Aono, James D. Love, Brandon Evans, Ronald D. Seidel, Brandan S. Hillerich, Scott J. Garforth, Steven C. Almo, Patrick S. Mariano, Debra Dunaway-Mariano, Karen N. Allen, Jeremiah D. Farelli

Research output: Contribution to journalArticle

56 Scopus citations

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 isofunctional 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.

Original languageEnglish (US)
Pages (from-to)E1974-E1983
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number16
DOIs
StatePublished - Apr 21 2015

Keywords

  • Evolution
  • Phosphatase
  • Promiscuity
  • Specificity
  • Substrate screen

ASJC Scopus subject areas

  • General

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    Huang, H., Pandya, C., Liu, C., Al-Obaidi, N. F., Wang, M., Zheng, L., Keating, S. T., Aono, M., Love, J. D., Evans, B., Seidel, R. D., Hillerich, B. S., Garforth, S. J., Almo, S. C., Mariano, P. S., Dunaway-Mariano, D., Allen, K. N., & Farelli, J. D. (2015). Panoramic view of a superfamily of phosphatases through substrate profiling. Proceedings of the National Academy of Sciences of the United States of America, 112(16), E1974-E1983. https://doi.org/10.1073/pnas.1423570112