期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 423, 期 4, 页码 555-575出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2012.08.017
关键词
ABC transporters; lignin degradation; Rhodopseudomonas palustris; solute-binding protein; benzoate
资金
- U.S. Department of Energy, Office of Biological and Environmental Research (BER), as part of BER's Genomic Science Program
- National Institutes of Health [GM094585]
- U.S. Department of Energy, BER [DE-AC02-06CH11357]
In vitro growth experiments have demonstrated that aromatic compounds derived from lignin can be metabolized and represent a major carbon resource for many soil bacteria. However, the proteins that mediate the movement of these metabolites across the cell membrane have not been thoroughly characterized. To address this deficiency, we used a library representative of lignin degradation products and a thermal stability screen to determine ligand specificity for a set of solute-binding proteins (SBPs) from ATP-binding cassette (ABC) transporters. The ligand mapping process identified a set of proteins from Alphaproteobacteria that recognize various benzoate derivatives. Seven high-resolution crystal structures of these proteins in complex with four different aromatic compounds were obtained. The protein ligand complexes provide details of molecular recognition that can be used to infer binding specificity. This structure function characterization provides new insight for the biological roles of these ABC transporters and their SBPs, which had been previously annotated as branched-chain amino-acid-binding proteins. The knowledge derived from the crystal structures provides a foundation for development of sequence-based methods to predict the ligand specificity of other uncharacterized transporters. These results also demonstrate that Alphaproteobacteria possess a diverse set of transport capabilities for lignin-derived compounds. Characterization of this new class of transporters improves genomic annotation projects and provides insight into the metabolic potential of soil bacteria. (C) 2012 Elsevier Ltd. All rights reserved.
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