title: Detection of glycoprotein receptors on blotting membranes by binding of live bacteria and amplification by growth.
abstract: Conditions have been adapted for detecting bacteria bound to glycoprotein receptors on blotting membranes using a self-enhancing detection method based on bacterial growth. Neutrophil plasma membrane proteins, mediating adherence of mannose-binding type-1-fimbriated Escherichia coli and concanavalin A (Con A) to intact human neutrophils, were separated by SDS-PAGE and transferred onto a polyvinylidene difluoride (PVDF) membrane. The PVDF membrane was immersed in a suspension of mannose-binding type-1-fimbriated E. coli, and after repeated washings, bound bacteria were allowed to multiply into bacterial colonies by placing the membrane on a solid nutrient substratum. About one major and eight minor glycoproteins, some of which also were detected by Con A, selectively induced colony formation in a mannose-inhibitable fashion. Binding of [35S]methionine metabolically labeled E. coli to PVDF membranes produced a virtually identical binding pattern, demonstrating further the accuracy of this self-enhancing detection method which is rapid, simple, and sensitive and avoids radioisotopes.
authors.affiliation: Phagocyte Research Laboratory, Department of Medical Microbiology and Immunology, Göteborg, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Karlsson
authors.forename: M
authors.initials: M
authors.lastname: Markfjäll
authors.forename: H
authors.initials: H
authors.lastname: Lundqvist
authors.forename: N
authors.initials: N
authors.lastname: Strömberg
authors.forename: C
authors.initials: C
authors.lastname: Dahlgren
mesh_terms: D001422:Bacterial Adhesion; D003208:Concanavalin A / Q000378:metabolism; D004926:Escherichia coli / Q000254:growth & development; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D009504:Neutrophils / Q000382:microbiology; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011145:Polyvinyls; D011956:Receptors, Cell Surface / Q000032:analysis*; D005465:Fluorocarbon Polymers
chemical_list: D003208:Concanavalin A; D008562:Membrane Glycoproteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011145:Polyvinyls; D011956:Receptors, Cell Surface; C032193:glycoprotein receptor GPIb-IX; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers