Congo Red Interactions with Curli-Producing E. coli and Native Curli Amyloid Fibers.
Microorganisms produce functional amyloids that can be examined and manipulated in vivo and in vitro. Escherichia coli assemble extracellular adhesive amyloid fibers termed curli that mediate adhesion and promote biofilm formation. We have characterized the dye binding properties of the hallmark amyloid dye, Congo red, with curliated E. coli and with isolated curli fibers. Congo red binds to curliated whole cells, does not inhibit growth, and can be used to comparatively quantify whole-cell curliation. Using Surface Plasmon Resonance, we measured the binding and dissociation kinetics of Congo red to curli. Furthermore, we determined that the binding of Congo red to curli is pH-dependent and that histidine residues in the CsgA protein do not influence Congo red binding. Our results on E. coli strain MC4100, the most commonly employed strain for studies of E. coli amyloid biogenesis, provide a starting point from which to compare the influence of Congo red binding in other E. coli strains and amyloid-producing organisms.
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Courtney
C
Reichhardt
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Amy N
AN
Jacobson
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Marie C
MC
Maher
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Jeremy
J
Uang
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Oscar A
OA
McCrate
Protein and Nucleic Acid Facility, Stanford University, Stanford, California 94305, United States of America.
Michael
M
Eckart
Department of Chemistry, Stanford University, Stanford, California 94305, United States of America.
Lynette
L
Cegelski
D000682:Amyloid / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D015153:Blotting, Western; D003224:Congo Red / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D011485:Protein Binding; D020349:Surface Plasmon Resonance
D000682:Amyloid; D001426:Bacterial Proteins; C077481:Crl protein, Bacteria; D003224:Congo Red