title: A 56 kDa binding protein for Escherichia coli heat-stable enterotoxin isolated from the cytoskeleton of rat intestinal membrane does not possess guanylate cyclase activity.
abstract: Proteins binding Escherichia coli heat-stable enterotoxin were isolated from the cytoskeleton of intestinal membranes using an affinity matrix of biotinylated ST immobilized on monomeric avidin-agarose. ST binding proteins were purified 343-fold using this affinity technique, with 7% of the initial binding activity recovered in these preparations. ST binding proteins isolated by affinity chromatography possessed a native and subunit molecular mass of 56 kDa. These preparations exhibited both high- and low-affinity binding sites for ST. Guanylate cyclase in extracts of the intestinal membrane cytoskeleton was completely recovered in fractions which did not associate with the affinity matrix. In addition, ST binding proteins isolated by affinity chromatography were devoid of guanylate cyclase activity. These data, taken together with those obtained previously with crude and partially purified receptors, suggest that ST binds to different proteins in intestinal membranes, some of which do not possess guanylate cyclase activity.
authors.affiliation: Department of Medicine, Thomas Jefferson University, Philadelphia, PA 19107.
authors.forename: S
authors.initials: S
authors.lastname: Hakki
authors.forename: D C
authors.initials: DC
authors.lastname: Robertson
authors.forename: S A
authors.initials: SA
authors.lastname: Waldman
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003599:Cytoskeleton / Q000737:chemistry; D004768:Enterotoxins / Q000378:metabolism*; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006162:Guanylate Cyclase / Q000032:analysis*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007421:Intestine, Small / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled; D018000:Receptors, Peptide / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D018000:Receptors, Peptide; C034151:heat stable toxin (E coli); D006162:Guanylate Cyclase; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled