Lysinoalanine crosslinking in the extracellular flagellar hook of Synergistota
- 1Cornel University
Abstract
Bacterial motility is driven by the flagellar motor, a conserved macromolecular complex that enables cells to navigate chemical gradients. In spirochetes, motility depends on periplasmic flagella whose function is supported by a unique post-translational modification (PTM) in the hook protein FlgE: a covalent lysinoalanine (Lal) crosslink between adjacent subunits. This modification has been shown to be conserved across the phylum and is required for motility. In spirochetes, Lal is thought to provide additional hook stability needed to meet the demands of flagellar rotation in the confined space of the periplasm and associated load of the wrapped cell body. Guided by a bioinformatic screen based on conserved catalytic residues required for Lal formation, we investigated the possibility that bacteria with extracellular flagella also produce Lal. Using a combination of biochemical crosslinking assays and high-resolution mass spectrometry, we demonstrate that FlgE from Thermanaerovibrio acidaminovorans, a motile member of the Synergistota phylum, forms Lal crosslinks analogous to those of spirochetes. These results establish the first example of Lal crosslinking in extracellular flagellar hooks and the first instance outside of the spirochete phylum. Thus, Lal crosslinking is more broadly conserved than previously appreciated and may represent a general mechanism for enhancing hook stability and function for extracellular as well as periplasmic flagella.
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BibTeX
@misc{lynch2026lysinoalanine,
title = {{Lysinoalanine crosslinking in the extracellular flagellar hook of Synergistota}},
author = {M. J. Lynch and N. W. Charon and B. R. Crane},
howpublished = {bioRxiv},
year = {2026},
month = may,
doi = {10.64898/2026.05.22.727228},
url = {https://www.biorxiv.org/content/10.64898/2026.05.22.727228v3},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - Lysinoalanine crosslinking in the extracellular flagellar hook of Synergistota
AU - M. J. Lynch
AU - N. W. Charon
AU - B. R. Crane
PY - 2026
DA - 2026/05/22
PB - bioRxiv
DO - 10.64898/2026.05.22.727228
UR - https://www.biorxiv.org/content/10.64898/2026.05.22.727228v3
AB - Bacterial motility is driven by the flagellar motor, a conserved macromolecular complex that enables cells to navigate chemical gradients. In spirochetes, motility depends on periplasmic flagella whose function is supported by a unique post-translational modification (PTM) in the hook protein FlgE: a covalent lysinoalanine (Lal) crosslink between adjacent subunits. This modification has been shown to be conserved across the phylum and is required for motility. In spirochetes, Lal is thought to provide additional hook stability needed to meet the demands of flagellar rotation in the confined space of the periplasm and associated load of the wrapped cell body. Guided by a bioinformatic screen based on conserved catalytic residues required for Lal formation, we investigated the possibility that bacteria with extracellular flagella also produce Lal. Using a combination of biochemical crosslinking assays and high-resolution mass spectrometry, we demonstrate that FlgE from Thermanaerovibrio acidaminovorans, a motile member of the Synergistota phylum, forms Lal crosslinks analogous to those of spirochetes. These results establish the first example of Lal crosslinking in extracellular flagellar hooks and the first instance outside of the spirochete phylum. Thus, Lal crosslinking is more broadly conserved than previously appreciated and may represent a general mechanism for enhancing hook stability and function for extracellular as well as periplasmic flagella.
ER -