title: Panoramic view of a superfamily of phosphatases through substrate profiling.
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.
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Hua
authors.initials: H
authors.lastname: Huang
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215; Bioinformatics Graduate Program, Boston University, Boston, MA 02215; and.
authors.forename: Chetanya
authors.initials: C
authors.lastname: Pandya
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Chunliang
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Nawar F
authors.initials: NF
authors.lastname: Al-Obaidi
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Li
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Sarah
authors.initials: S
authors.lastname: Toews Keating
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215;
authors.forename: Miyuki
authors.initials: M
authors.lastname: Aono
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: James D
authors.initials: JD
authors.lastname: Love
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Brandon
authors.initials: B
authors.lastname: Evans
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Seidel
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Brandan S
authors.initials: BS
authors.lastname: Hillerich
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Garforth
authors.affiliation: Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Almo
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Mariano
authors.affiliation: Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, NM 87131;
authors.forename: Debra
authors.initials: D
authors.lastname: Dunaway-Mariano
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215; drkallen@bu.edu jfarelli@bu.edu.
authors.forename: Karen N
authors.initials: KN
authors.lastname: Allen
authors.affiliation: Department of Chemistry, Boston University, Boston, MA 02215; drkallen@bu.edu jfarelli@bu.edu.
authors.forename: Jeremiah D
authors.initials: JD
authors.lastname: Farelli
mesh_terms: D057166:High-Throughput Screening Assays; D007700:Kinetics; D005810:Multigene Family; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D015203:Reproducibility of Results; D013379:Substrate Specificity
chemical_list: D010744:Phosphoric Monoester Hydrolases
keywords: evolution; phosphatase; promiscuity; specificity; substrate screen
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