Inhibition of P-fimbriated Escherichia coli adhesion by multivalent galabiose derivatives studied by a live-bacteria application of surface plasmon resonance.
OBJECTIVE
Uropathogenic P-fimbriated Escherichia coli adheres to host cells by specific adhesins recognizing galabiose (Galalpha1-4Gal)-containing structures on cell surfaces. In search of agents inhibiting this first step of infection, the inhibition potency of a set of synthetic mono- and multivalent galabiose compounds was evaluated. In order to mimic the flow conditions of natural infections, a live-bacteria application of surface plasmon resonance (SPR) was established.


RESULTS
For the measurement of the binding of E. coli to a surface containing galabiose, live bacteria were injected over the flow cell, and the inhibition of adhesion caused by the galabiose inhibitors was recorded. Quantitative binding data were recorded in real-time for each inhibitor. The results were compared with those of conventional static haemagglutination and ELISA-based cell adhesion assays. Compared with the Gram-positive Streptococcus suis bacteria, which also bind to galabiose and whose binding inhibition is strongly dependent on the multivalency of the inhibitor, E. coli inhibition was only moderately affected by the valency. However, a novel octavalent compound was found to be the most effective inhibitor of E. coli PapG(J96) adhesion, with an IC50 value of 2 microM.


CONCLUSIONS
Measurement of bacterial adhesion by SPR is an efficient way to characterize the adhesion of whole bacterial cells and allows the characterization of the inhibitory potency of adhesion inhibitors under dynamic flow conditions. Under these conditions, multivalency increases the anti-adhesion potency of galabiose-based inhibitors of P-fimbriated E. coli adhesion and provides a promising approach for the design of high-affinity anti-adhesion agents.
Salminen
Annika
A
Department of Medical Biochemistry and Molecular Biology, University of Turku, FI-20520 Turku, Finland.
Loimaranta
Vuokko
V
Joosten
John A F
JA
Khan
A Salam
AS
Hacker
Jörg
J
Pieters
Roland J
RJ
Finne
Jukka
J
D001422:Bacterial Adhesion / Q000187:drug effects*; D002241:Carbohydrates / Q000737:chemistry; D004187:Disaccharides / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D010861:Fimbriae, Bacterial / Q000502:physiology*; D006386:Hemagglutination Tests; D012710:Serum Albumin, Bovine / Q000737:chemistry; D017011:Streptococcus suis / Q000187:drug effects; D020349:Surface Plasmon Resonance
D002241:Carbohydrates; D004187:Disaccharides; D012710:Serum Albumin, Bovine; C056853:6-O-galactopyranosylgalactose