期刊
CHEMBIOCHEM
卷 20, 期 18, 页码 2360-2372出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201900184
关键词
cytochrome c; lanthanides; methanol dehydrogenase; rare earths; reduction potential
资金
- DOE Office of Science [DE-AC02-06CH11357]
- National Cancer Institute [ACB-12002]
- National Institute of General Medical Sciences (NIGMS) [AGM-12006]
- NIH Office of Research Infrastructure Programs, High-End Instrumentation grant [1S10OD012289-01A1]
- Michigan Economic Development Corporation
- Michigan Technology Tri-Corridor Grant [085P1000817]
- NIH, NIGMS
- Howard Hughes Medical Institute
- DOE Office of Science User Facility [DE-AC02-05CH11231]
- Huck Institutes for the Life Sciences
- Louis Martarano Career Development Professorship
- NIH [GM119707]
- Penn State Department of Chemistry
Lanthanide (Ln)-dependent methanol dehydrogenases (MDHs) have recently been shown to be widespread in methylotrophic bacteria. Along with the core MDH protein, XoxF, these systems contain two other proteins, XoxG (a c-type cytochrome) and XoxJ (a periplasmic binding protein of unknown function), about which little is known. In this work, we have biochemically and structurally characterized these proteins from the methyltroph Methylobacterium extorquens AM1. In contrast to results obtained in an artificial assay system, assays of XoxFs metallated with La-III, Ce-III, and Nd-III using their physiological electron acceptor, XoxG, display Ln-independent activities, but the K-m for XoxG markedly increases from La to Nd. This result suggests that XoxG ' s redox properties are tuned specifically for lighter Lns in XoxF, an interpretation supported by the unusually low reduction potential of XoxG (+172 mV). The X-ray crystal structure of XoxG provides a structural basis for this reduction potential and insight into the XoxG-XoxF interaction. Finally, the X-ray crystal structure of XoxJ reveals a large hydrophobic cleft and suggests a role in the activation of XoxF. These studies enrich our understanding of the underlying chemical principles that enable the activity of XoxF with multiple lanthanides in vitro and in vivo.
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