Spirochaete flagella hook proteins self-catalyse a lysinoalanine covalent crosslink for motility.
Spirochaetes are bacteria responsible for several serious diseases, including Lyme disease (Borrelia burgdorferi), syphilis (Treponema pallidum) and leptospirosis (Leptospira interrogans), and contribute to periodontal diseases (Treponema denticola)(1). These spirochaetes employ an unusual form of flagella-based motility necessary for pathogenicity; indeed, spirochaete flagella (periplasmic flagella) reside and rotate within the periplasmic space(2-11). The universal joint or hook that links the rotary motor to the filament is composed of ∼120-130 FlgE proteins, which in spirochaetes form an unusually stable, high-molecular-weight complex(9,12-17). In other bacteria, the hook can be readily dissociated by treatments such as heat(18). In contrast, spirochaete hooks are resistant to these treatments, and several lines of evidence indicate that the high-molecular-weight complex is the consequence of covalent crosslinking(12,13,17). Here, we show that T. denticola FlgE self-catalyses an interpeptide crosslinking reaction between conserved lysine and cysteine, resulting in the formation of an unusual lysinoalanine adduct that polymerizes the hook subunits. Lysinoalanine crosslinks are not needed for flagellar assembly, but they are required for cell motility and hence infection. The self-catalytic nature of FlgE crosslinking has important implications for protein engineering, and its sensitivity to chemical inhibitors provides a new avenue for the development of antimicrobials targeting spirochaetes.
Department of Biochemistry, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Michael R
MR
Miller
Department of Microbiology, Immunology, and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Kelly A
KA
Miller
Department of Oral Biology, State University of New York, Buffalo, New York 14214, USA.
Jiang
J
Bian
Department of Microbiology, Immunology, and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Milinda E
ME
James
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Sheng
S
Zhang
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Michael J
MJ
Lynch
Department of Pharmaceutical Sciences, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Patrick S
PS
Callery
National Institute for Occupational Safety and Health, 1095 Willowdale Road, Morgantown West Virginia 26505, USA.
Justin M
JM
Hettick
Department of Microbiology, Immunology, and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Andrew
A
Cockburn
Department of Pathology and Laboratory Medicine, University of Texas Health Sciences Center, Houston, Texas 77030, USA.
Jun
J
Liu
Department of Oral Biology, State University of New York, Buffalo, New York 14214, USA.
Chunhao
C
Li
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Brian R
BR
Crane
Department of Microbiology, Immunology, and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, USA.
Nyles W
NW
Charon
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D055162:Biocatalysis; D025065:Borrelia burgdorferi / Q000378:metabolism; D005407:Flagella / Q000737:chemistry* / Q000502:physiology; D008241:Lysinoalanine / Q000737:chemistry / Q000378:metabolism*; D009068:Movement; D013142:Spirochaeta / Q000378:metabolism* / Q000472:pathogenicity; D048373:Treponema denticola / Q000378:metabolism
D001426:Bacterial Proteins; C035832:hook protein, bacterial flagellum; D008241:Lysinoalanine
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