Abstract
Methanotrophic bacteria use methane, a potent greenhouse gas, as their primary source of carbon and energy. The first step in methane metabolism is its oxidation to methanol. In almost all methanotrophs, this chemically challenging reaction is catalyzed by particulate methane monooxygenase (pMMO), a copper-dependent integral membrane enzyme. Methanotrophs acquire copper (Cu) for pMMO by secreting a small ribosomally produced, posttranslationally modified natural product called methanobactin (Mbn). Mbn chelates Cu with high affinity, and the Cu-loaded form (CuMbn) is reinternalized into the cell via an active transport process. Bioinformatic and gene regulation studies suggest that two proteins might play a role in CuMbn handling: the TonB-dependent transporter MbnT and the periplasmic binding protein MbnE. Disruption of the gene that encodes MbnT abolishes CuMbn uptake, as reported previously, and expression of MbnT in Escherichia coli confers the ability to take up CuMbn. Biophysical studies of MbnT and MbnE reveal specific interactions with CuMbn, and a crystal structure of apo MbnE is consistent with MbnE's proposed role as a periplasmic CuMbn transporter. Notably, MbnT and MbnE exhibit different levels of discrimination between cognate and noncognate CuMbns. These findings provide evidence for CuMbn-protein interactions and begin to elucidate the molecular mechanisms of its recognition and transport.
Original language | English (US) |
---|---|
Pages (from-to) | 13027-13032 |
Number of pages | 6 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 113 |
Issue number | 46 |
DOIs | |
State | Published - Nov 15 2016 |
Funding
This work was provided by National Science Foundation Grant MCB0842366 (to A.C.R.) and National Institutes of Health Grant GM118035 (to A.C.R.) and Grant F32GM110934 (to L.M.K.D.). L.M.K.D. is the recipient of a Postdoctoral Enrichment Program grant from the Burroughs Wellcome Fund. G.E.K. was supported by American Heart Association Predoctoral Fellowship 14PRE20460104. Constructs of the Ms. trichosporium OB3b and Mc. parvus OBBP MbnEs were provided by Anthony S. Gizzi and Steven C. Almo (Albert Einstein College of Medicine), who are supported by the Price Family Foundation and New York Structural Genomics Research Center (National Institute of General Medical Sciences Grant U54-GM094663). Additional institutional and core facility acknowledgments are in the SI Appendix.
Keywords
- Chalkophore
- Copper transport
- Metal homeostasis
- Methane monooxygenase
- Methanobactin
ASJC Scopus subject areas
- General
Fingerprint
Dive into the research topics of 'Methanobactin transport machinery'. Together they form a unique fingerprint.Datasets
-
Methanobactin periplasmic binding protein
Dassama, L. M. K. (Contributor), Kenney, G. E. (Contributor), Ro, S. Y. (Contributor), Zielazinski, E. L. (Contributor) & Rosenzweig, A. C. (Contributor), Protein Data Bank (PDB), Oct 26 2016
DOI: 10.2210/pdb5ICQ/pdb, https://www.wwpdb.org/pdb?id=pdb_00005icq
Dataset