Role of the intramolecular disulfide bond in FlgI, the flagellar P-ring component of Escherichia coli.
The P ring of the bacterial flagellar motor consists of multiple copies of FlgI, a periplasmic protein. The intramolecular disulfide bond in FlgI has previously been reported to be essential for P-ring assembly in Escherichia coli, because the P ring was not assembled in a dsbB strain that was defective for disulfide bond formation in periplasmic proteins. We, however, found that the two Cys residues of FlgI are not conserved in other bacterial species. We then assessed the role of this intramolecular disulfide bond in FlgI. A Cys-eliminated FlgI derivative formed a P ring that complemented the flagellation defect of our DeltaflgI strain when it was overproduced, suggesting that disulfide bond formation in FlgI is not absolutely required for P-ring assembly. The levels of the mature forms of the FlgI derivatives were significantly lower than that of wild-type FlgI, although the precursor protein levels were unchanged. Moreover, the FlgI derivatives were more susceptible to degradation than wild-type FlgI. Overproduction of FlgI suppressed the motility defect of DeltadsbB cells. Additionally, the low level of FlgI observed in the DeltadsbB strain increased in the presence of l-cystine, an oxidative agent. We propose that intramolecular disulfide bond formation facilitates the rapid folding of the FlgI monomer to protect against degradation in the periplasmic space, thereby allowing its efficient self-assembly into the P ring.
Hizukuri
Yohei
Y
Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 8304 Minamiminowa, Nagano 399-4598, Japan.
Yakushi
Toshiharu
T
Kawagishi
Ikuro
I
Homma
Michio
M
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D003553:Cystine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism / Q000502:physiology*; D005407:Flagella / Q000737:chemistry / Q000378:metabolism*; D008124:Locomotion; D017510:Protein Folding
D001426:Bacterial Proteins; D004220:Disulfides; C074899:flgI protein, Bacteria; D003553:Cystine