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Stewart, A.* ; Dershwitz, P.* ; Stewart, C.* ; Sawaya, M.R.* ; Yeates, T.O.* ; Semrau, J.D.* ; Zischka, H. ; DiSpirito, A.A.* ; Bobik, T.A.*

Crystal structure of MbnF: An NADPH-dependent flavin monooxygenase from Methylocystis strain SB2.

Acta Crystallogr. F-Struct. Biol. Cryst. Commun. 79, 111-118 (2023)
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Methanobactins (MBs) are ribosomally produced and post-translationally modified peptides (RiPPs) that are used by methanotrophs for copper acquisition. The signature post-translational modification of MBs is the formation of two heterocyclic groups, either an oxazolone, pyrazinedione or imidazolone group, with an associated thioamide from an X-Cys dipeptide. The precursor peptide (MbnA) for MB formation is found in a gene cluster of MB-associated genes. The exact biosynthetic pathway of MB formation is not yet fully understood, and there are still uncharacterized proteins in some MB gene clusters, particularly those that produce pyrazinedione or imidazolone rings. One such protein is MbnF, which is proposed to be a flavin monooxygenase (FMO) based on homology. To help to elucidate its possible function, MbnF from Methylocystis sp. strain SB2 was recombinantly produced in Escherichia coli and its X-ray crystal structure was resolved to 2.6 Å resolution. Based on its structural features, MbnF appears to be a type A FMO, most of which catalyze hydroxylation reactions. Preliminary functional characterization shows that MbnF preferentially oxidizes NADPH over NADH, supporting NAD(P)H-mediated flavin reduction, which is the initial step in the reaction cycle of several type A FMO enzymes. It is also shown that MbnF binds the precursor peptide for MB, with subsequent loss of the leader peptide sequence as well as the last three C-terminal amino acids, suggesting that MbnF might be needed for this process to occur. Finally, molecular-dynamics simulations revealed a channel in MbnF that is capable of accommodating the core MbnA fragment minus the three C-terminal amino acids.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Mbnf ; Methylocystis Sp. Strain Sb2 ; Wilson's Disease ; Flavin Monooxygenases ; Methanobactins; Methanobactin; Copper; Binding; Mercury
e-ISSN 2053-230X
Journal Acta Crystallographica Section F
Quellenangaben Volume: 79, Issue: Pt 5, Pages: 111-118 Article Number: , Supplement: ,
Publisher Blackwell
Publishing Place Oxford [u.a.]
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Molecular Toxicology and Pharmacology (TOXI)
Grants U.S. Department of Energy (DOE)
ISU Bailey Research and Career Development Award
National Science Foundation
US Department of Energy Office of Science