A Defined and Flexible Pocket Explains Aryl Substrate Promiscuity of the Cahuitamycin Starter Unit-Activating Enzyme CahJ

Ashootosh Tripathi; Sung Ryeol Park; Andrew P Sikkema; Hyo Je Cho; Jianfeng Wu; Brian Lee; Chuanwu Xi; Janet L Smith; David H Sherman

doi:10.1002/cbic.201800233

A Defined and Flexible Pocket Explains Aryl Substrate Promiscuity of the Cahuitamycin Starter Unit-Activating Enzyme CahJ

Chembiochem. 2018 Aug 6;19(15):1595-1600. doi: 10.1002/cbic.201800233. Epub 2018 Jun 21.

Authors

Ashootosh Tripathi^{1

2}, Sung Ryeol Park^{1

3}, Andrew P Sikkema^{1

4

5}, Hyo Je Cho⁶, Jianfeng Wu⁷, Brian Lee¹, Chuanwu Xi⁷, Janet L Smith^{1

4}, David H Sherman^{1

2

8

9}

Affiliations

¹ Life Sciences Institute, University of Michigan, 210 Washtenaw Avenue, Ann Arbor, MI, 48109-2216, USA.
² Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
³ Baruch S. Blumberg Institute, Natural Products Discovery Institute, 3805 Old Easton Road, Doylestown, PA, 18902, USA.
⁴ Department of Biological Chemistry, University of Michigan, 1150 W. Medical Center Drive, Ann Arbor, MI, 48109, USA.
⁵ Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, 27709, USA.
⁶ Department of Pathology, University of Michigan, Ann Arbor, MI, 48109, USA.
⁷ Department of Environmental Health Sciences, University of Michigan School of Public Health, Ann Arbor, MI, 48109, USA.
⁸ Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
⁹ Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109, USA.

Abstract

Cahuitamycins are biofilm inhibitors assembled by a convergent nonribosomal peptide synthetase pathway. Previous genetic analysis indicated that a discrete enzyme, CahJ, serves as a gatekeeper for cahuitamycin structural diversification. Here, the CahJ protein was probed structurally and functionally to guide the formation of new analogues by mutasynthetic studies. This analysis enabled the in vivo production of a new cahuitamycin congener through targeted precursor incorporation.

Keywords: adenylation domains; biosynthesis; kinetics; natural products; protein structures.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / chemistry
Bacterial Proteins / metabolism*
Binding Sites
Biosynthetic Pathways
Molecular Docking Simulation
Oligopeptides / chemistry
Oligopeptides / metabolism*
Peptide Synthases / chemistry
Peptide Synthases / metabolism*
Protein Conformation
Streptomyces / chemistry
Streptomyces / metabolism*
Substrate Specificity

Substances

Bacterial Proteins
Oligopeptides
cahuitamycin A protein, Streptomyces gandocaensis
cahuitamycin B protein, Streptomyces gandocaensis
cahuitamycin C protein, Streptomyces gandocaensis
Peptide Synthases
non-ribosomal peptide synthase

Abstract

Publication types

MeSH terms

Substances

Grants and funding