Structural and spectroscopic analyses of the sporulation killing factor biosynthetic enzyme SkfB, a bacterial AdoMet radical sactisynthase

Grell, Tsehai A.J.; Kincannon, William M.; Bruender, Nathan A.; Blaesi, Elizabeth J.; Krebs, Carsten; Bandarian, Vahe; Drennan, Catherine L.

doi:10.1074/jbc.ra118.005369

Title: Structural and spectroscopic analyses of the sporulation killing factor biosynthetic enzyme SkfB, a bacterial AdoMet radical sactisynthase

Journal Article · Fri Sep 14 00:00:00 EDT 2018 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.ra118.005369· OSTI ID:1570710

Grell, Tsehai A.J. ^[1]; Kincannon, William M. ^[2]; Bruender, Nathan A. ^[2]; Blaesi, Elizabeth J. ^[3]; Krebs, Carsten ^[3]; Bandarian, Vahe ^[2]; Drennan, Catherine L. ^[1]

Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
Univ. of Utah, Salt Lake City, UT (United States)
Pennsylvania State Univ., University Park, PA (United States)

Sactipeptides are a subclass of ribosomally synthesized and post-translationally modified peptides (RiPPs). They contain a unique thioether bond, referred to as a sactionine linkage, between the sulfur atom of a cysteine residue and the α-carbon of an acceptor residue. These linkages are formed via radical chemistry and are essential for the spermicidal, antifungal, and antibacterial properties of sactipeptides. Enzymes that form these linkages, called sactisynthases, are AdoMet radical enzymes in the SPASM/Twitch subgroup whose structures are incompletely characterized. Here, we present the X-ray crystal structure to 1.29-Å resolution and Mössbauer analysis of SkfB, a sactisynthase from Bacillus subtilis involved in making sporulation killing factor (SKF). We found that SkfB is a modular enzyme with an N-terminal peptide-binding domain comprising a RiPP recognition element (RRE), a middle domain that forms a classic AdoMet radical partial (β/α)₆ barrel structure and displays AdoMet bound to the [4Fe-4S] cluster, and a C-terminal region characteristic of the so-called Twitch domain housing an auxiliary iron-sulfur cluster. Notably, both crystallography and Mössbauer analyses suggest that SkfB can bind a [2Fe-2S] cluster at the auxiliary cluster site, which has been observed only once before in a SPASM/Twitch auxiliary cluster site in the structure of another AdoMet radical enzyme, the pyrroloquinoline quinone biosynthesis enzyme PqqE. Taken together, our findings indicate that SkfB from B. subtilis represents a unique enzyme containing several structural features observed in other AdoMet radical enzymes.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC)National Institutes of Health (NIH)

Grant/Contract Number:: AC02-06CH11357; P41 GM103403; S10 RR029205

OSTI ID:: 1570710

Journal Information:: Journal of Biological Chemistry, Vol. 293, Issue 45; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 23 works

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Title: Structural and spectroscopic analyses of the sporulation killing factor biosynthetic enzyme SkfB, a bacterial AdoMet radical sactisynthase

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