Binding proteins for cyclosomatostatins and bile acids in basolateral plasma membranes of rat liver.
The bile acids cholate and taurocholate on the one hand and the cyclopeptide c(Phe-Thr-Lys-Trp-Phe-D-Pro) (008), an analog of somatostatin with retro sequence, on the other hand, display mutually competitive transport inhibition into isolated rat hepatocytes. This indicates a common transport system for bile acids and cyclosomatostatins in sinusoidal rat liver plasma membranes. In order to identify and isolate common binding and/or transport proteins for bile acids and the cyclopeptides by affinity chromatography, the bile acid derivative 4'-amino-7-benzamidotaurocholate (ABATC) and the cyclosomatostatin-analog 008 were attached to a gel matrix. Two methods were used to prepare integral membrane proteins: (1) alkaline EDTA extraction and (2) Triton X-114 phase separation. Octyl glycoside solubilized, alkaline EDTA-extracted integral basolateral membrane proteins with apparent molecular masses of 52 and 48 kDa bound specifically to the ABATC affinity matrix. Two-phase Triton X-114 separated integral membrane proteins of the same molecular masses bound specifically to the cyclosomatostatin ligand. The 48 kDa ABATC and 008 binding protein was shown to be present in the basolateral plasma membrane fraction and in the microsomal fraction. The isolated 52 kDa ABATC binding protein was localized only in basolateral plasma membranes and could not be found in isolated microsomes.
Institut für Pharmakologie und Toxikologie der Justus-Liebig-Universität, Giessen, Germany.
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Wenzel
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Ziegler
D000595:Amino Acid Sequence; D000818:Animals; D001386:Azides / Q000378:metabolism; D001647:Bile Acids and Salts / Q000378:metabolism*; D002352:Carrier Proteins / Q000032:analysis*; D002462:Cell Membrane / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013656:Taurocholic Acid / Q000031:analogs & derivatives / Q000378:metabolism
D001386:Azides; D001647:Bile Acids and Salts; D002352:Carrier Proteins; D010456:Peptides, Cyclic; C067675:cyclosomatostatin; C059201:azidobenzamidotaurocholate; D013656:Taurocholic Acid