The cysteine bond in the Escherichia coli FimH adhesin is critical for adhesion under flow conditions.
Cysteine bonds are found near the ligand-binding sites of a wide range of microbial adhesive proteins, including the FimH adhesin of Escherichia coli. We show here that removal of the cysteine bond in the mannose-binding domain of FimH did not affect FimH-mannose binding under static or low shear conditions (< or = 0.2 dyne cm(-2)). However, the adhesion level was substantially decreased under increased fluid flow. Under intermediate shear (2 dynes cm(-2)), the ON-rate of bacterial attachment was significantly decreased for disulphide-free mutants. Molecular dynamics simulations demonstrated that the lower ON-rate of cysteine bond-free FimH could be due to destabilization of the mannose-free binding pocket of FimH. In contrast, mutant and wild-type FimH had similar conformation when bound to mannose, explaining their similar binding strength to mannose under intermediate shear. The stabilizing effect of mannose on disulphide-free FimH was also confirmed by protection of the FimH from thermal and chemical inactivation in the presence of mannose. However, this stabilizing effect could not protect the integrity of FimH structure under high shear (> 20 dynes cm(-2)), where lack of the disulphide significantly increased adhesion OFF-rates. Thus, the cysteine bonds in bacterial adhesins could be adapted to enable bacteria to bind target surfaces under increased shear conditions.
Nilsson
Lina M
LM
Department of Materials, ETH Zurich, Hönggerberg, CH-8093 Zürich, Switzerland.
Yakovenko
Olga
O
Tchesnokova
Veronika
V
Thomas
Wendy E
WE
Schembri
Mark A
MA
Vogel
Viola
V
Klemm
Per
P
Sokurenko
Evgeni V
EV
D018830:Adhesins, Escherichia coli / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001422:Bacterial Adhesion; D001665:Binding Sites; D003545:Cysteine / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000737:chemistry / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D033081:Shear Strength
D018830:Adhesins, Escherichia coli; D004220:Disulfides; C058769:fimH protein, E coli; D036781:Fimbriae Proteins; D003545:Cysteine; D008358:Mannose