Structure and chemistry of lysinoalanine crosslinking in the spirochaete flagella hook

Lynch, Michael J.; Miller, Michael; James, Milinda; Zhang, Sheng; Zhang, Kai; Li, Chunhao; Charon, Nyles W.; Crane, Brian R.

doi:10.1038/s41589-019-0341-3

Structure and chemistry of lysinoalanine crosslinking in the spirochaete flagella hook

Journal Article · Mon Aug 12 00:00:00 EDT 2019 · Nature Chemical Biology

DOI:https://doi.org/10.1038/s41589-019-0341-3· OSTI ID:1569895

Lynch, Michael J. ^[1]; Miller, Michael ^[2]; James, Milinda ^[2]; Zhang, Sheng ^[1]; Zhang, Kai ^[3]; Li, Chunhao ^[3]; Charon, Nyles W. ^[2]; ^[1]

Cornell Univ., Ithaca, NY (United States)
West Virginia Univ., Morgantown, WV (United States)
Virginia Commonwealth Univ., Richmond, VA (United States)

The flagellar hook protein FlgE from spirochaete bacteria self-catalyzes the formation of an unusual inter-subunit lysinoalanine (Lal) crosslink that is critical for cell motility. Unlike other known examples of Lal biosynthesis, conserved cysteine and lysine residues in FlgE spontaneously react to form Lal without the involvement of additional enzymes. Oligomerization of FlgE via its D0 and Dc domains drives assembly of the crosslinking site at the D1–D2 domain interface. Here, structures of the FlgED2 domain, dehydroalanine (DHA) intermediate and Lal crosslinked FlgE subunits reveal successive snapshots of the reaction. Cys178 flips from a buried configuration to release hydrogen sulfide (H₂S/HS^–) and produce DHA. Interface residues provide hydrogen bonds to anchor the active site, facilitate β-elimination of Cys178 and polarize the peptide backbone to activate DHA for reaction with Lys165. Cysteine-reactive molecules accelerate DHA formation, whereas nucleophiles can intercept the DHA intermediate, thereby indicating a potential for Lal crosslink inhibitors to combat spirochaetal diseases.

View Accepted Manuscript (DOE)

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

Sponsoring Organization:: USDOE; National Institutes of Health (NIH); National Science Foundation (NSF)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1569895

Journal Information:: Nature Chemical Biology, Journal Name: Nature Chemical Biology Journal Issue: 10 Vol. 15; ISSN 1552-4450

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Structure and chemistry of lysinoalanine crosslinking in the spirochaete flagella hook

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