title: Structure and Inhibition of Microbiome β-Glucuronidases Essential to the Alleviation of Cancer Drug Toxicity.
abstract: The selective inhibition of bacterial β-glucuronidases was recently shown to alleviate drug-induced gastrointestinal toxicity in mice, including the damage caused by the widely used anticancer drug irinotecan. Here, we report crystal structures of representative β-glucuronidases from the Firmicutes Streptococcus agalactiae and Clostridium perfringens and the Proteobacterium Escherichia coli, and the characterization of a β-glucuronidase from the Bacteroidetes Bacteroides fragilis. While largely similar in structure, these enzymes exhibit marked differences in catalytic properties and propensities for inhibition, indicating that the microbiome maintains functional diversity in orthologous enzymes. Small changes in the structure of designed inhibitors can induce significant conformational changes in the β-glucuronidase active site. Finally, we establish that β-glucuronidase inhibition does not alter the serum pharmacokinetics of irinotecan or its metabolites in mice. Together, the data presented advance our in vitro and in vivo understanding of the microbial β-glucuronidases, a promising new set of targets for controlling drug-induced gastrointestinal toxicity.
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Bret D
authors.initials: BD
authors.lastname: Wallace
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Adam B
authors.initials: AB
authors.lastname: Roberts
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Pollet
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: James D
authors.initials: JD
authors.lastname: Ingle
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Kristen A
authors.initials: KA
authors.lastname: Biernat
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Pellock
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Madhu Kumar
authors.initials: MK
authors.lastname: Venkatesh
authors.affiliation: Department of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Guthrie
authors.affiliation: Department of Pharmacotherapy and Experimental Therapeutics, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Sara K
authors.initials: SK
authors.lastname: O'Neal
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Sara J
authors.initials: SJ
authors.lastname: Robinson
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Makani
authors.initials: M
authors.lastname: Dollinger
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Esteban
authors.initials: E
authors.lastname: Figueroa
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Sarah R
authors.initials: SR
authors.lastname: McShane
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Rachel D
authors.initials: RD
authors.lastname: Cohen
authors.affiliation: Department of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
authors.affiliation: Department of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: Stephen V
authors.initials: SV
authors.lastname: Frye
authors.affiliation: Department of Pharmacotherapy and Experimental Therapeutics, University of North Carolina at Chapel Hill, NC 27599-3290, USA.
authors.forename: William C
authors.initials: WC
authors.lastname: Zamboni
authors.affiliation: Department of Soil and Crop Sciences, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Pepe-Ranney
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Sridhar
authors.initials: S
authors.lastname: Mani
authors.affiliation: Department of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Libusha
authors.initials: L
authors.lastname: Kelly
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, NC 27599-3290, USA; Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, NC 27599-3290, USA. Electronic address: redinbo@unc.edu.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Redinbo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D001441:Bacteroides fragilis / Q000201:enzymology; D002166:Camptothecin / Q000031:analogs & derivatives / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity; D003016:Clostridium perfringens / Q000201:enzymology; D004354:Drug Screening Assays, Antitumor; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D004926:Escherichia coli / Q000201:enzymology; D005966:Glucuronidase / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D000077146:Irinotecan; D051379:Mice; D008807:Mice, Inbred BALB C; D064307:Microbiota / Q000187:drug effects*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D013292:Streptococcus agalactiae / Q000201:enzymology
chemical_list: D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D000077146:Irinotecan; D005966:Glucuronidase; D002166:Camptothecin
keywords: NSAIDs; chemotherapy-induced diarrhea; irinotecan; microbiota