Structural and Functional Basis for Targeting Campylobacter jejuni Agmatine Deiminase to Overcome Antibiotic Resistance

Roger Shek, Devon A. Dattmore, Devin P. Stives, Ashley L. Jackson, Christa H. Chatfield, Katherine A. Hicks, Jarrod B. French

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Campylobacter jejuni is the most common bacterial cause of gastroenteritis and a major contributor to infant mortality in the developing world. The increasing incidence of antibiotic-resistant C. jejuni only adds to the urgency to develop effective therapies. Because of the essential role that polyamines play, particularly in protection from oxidative stress, enzymes involved in the biosynthesis of these metabolites are emerging as promising antibiotic targets. The recent description of an alternative pathway for polyamine synthesis, distinct from that in human cells, in C. jejuni suggests this pathway could be a target for novel therapies. To that end, we determined X-ray crystal structures of C. jejuni agmatine deiminase (CjADI) and demonstrated that loss of CjADI function contributes to antibiotic sensitivity, likely because of polyamine starvation. The structures provide details of key molecular features of the active site of this protein. Comparison of the unliganded structure (2.1 Å resolution) to that of the CjADI-agmatine complex (2.5 Å) reveals significant structural rearrangements that occur upon substrate binding. The shift of two helical regions of the protein and a large conformational change in a loop near the active site generate a narrow binding pocket around the bound substrate. This change optimally positions the substrate for catalysis. In addition, kinetic analysis of this enzyme demonstrates that CjADI is an iminohydrolase that effectively deiminates agmatine. Our data suggest that C. jejuni agmatine deiminase is a potentially important target for combatting antibiotic resistance, and these results provide a valuable framework for guiding future drug development.

Original languageEnglish (US)
Pages (from-to)6734-6742
Number of pages9
JournalBiochemistry
Volume56
Issue number51
DOIs
StatePublished - Dec 26 2017
Externally publishedYes

Bibliographical note

Funding Information:
*Phone: 607-753-2235. E-mail: christa.chatfield@cortland.edu. *Phone: 607-753-4324. E-mail: katherine.hicks@cortland.edu. *Phone: 631-632-8015. E-mail: jarrod.french@stonybrook.edu. ORCID Jarrod B. French: 0000-0002-6762-1309 Funding R.S. gratefully acknowledges support from the Biochemistry and Structural Biology Graduate Training Program at Stony Brook University, funded by National Institutes of Health Grant T32GM00846. Parts of this work were supported by the National Institute of General Medical Sciences of the National Institutes of Health under Grant R35GM124898 (J.B.F.) and the National Science Foundation under Grant 1337695 (C.H.C.). This work was also supported by funding from the SUNY Cortland Faculty Research Program (K.A.H. and C.H.C.). Notes The authors declare no competing financial interest.

Funding Information:
The authors thank Bruce M. Pearson for the kind gift of C. jejuni strain C8J_0892. This work is based upon research conducted at the Northeastern Collaborative Access Team (NE-CAT) facility at the Advanced Photon Source and the MacCHESS facility at the Cornell High Energy Synchrotron Source (CHESS). NE-CAT beamlines are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P41 GM103403). The Pilatus 6M detector on 24-ID-C beamline is funded by a NIH-ORIP HEI grant (S10 RR029205). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. CHESS is supported by the National Science Foundation (NSF) and the National Institutes of Health/National Institute of General Medical Sciences under NSF Grant DMR-0936384, and the Macromolecular Diffraction at CHESS (MacCHESS) facility is supported by National Institutes of Health Grant GM-103485, through its National Institute of General Medical Sciences.

Funding Information:
This work is based upon research conducted at the Northeastern Collaborative Access Team (NE-CAT) facility at the Advanced Photon Source and the MacCHESS facility at the Cornell High Energy Synchrotron Source (CHESS). NE-CAT beamlines are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P41 GM103403). The Pilatus 6M detector on 24-ID-C beamline is funded by a NIH-ORIP HEI grant (S10 RR029205). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. CHESS is supported by the National Science Foundation (NSF) and the National Institutes of Health/National Institute of General Medical Sciences under NSF Grant DMR-0936384, and the Macromolecular Diffraction at CHESS (MacCHESS) facility is supported by National Institutes of Health Grant GM-103485, through its National Institute of General Medical Sciences

Publisher Copyright:
© 2017 American Chemical Society.

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