Lithocholate binding by Y and Y' proteins in bovine small intestine.
Cytosolic proteins may play an important role in the intracellular transport of bile acids in enterocytes. The lithocholate binding properties of cytosolic protein from bovine small intestine were studied. Lithocholate binding was observed in the Y (45-50 kDa), Y' (30-35 kDa), and Z fractions (10-15 kDa) following gel filtration of cytosol. A Y protein with glutathione S-transferase activity (46 kDa) was purified by S-octyl-glutathione affinity chromatography and chromatofocusing (eluted at pH 7.5) of the Y fraction. Two Y' bile acid binding proteins with dihydrodiol dehydrogenase activity were partially purified from the Y' fraction by chromatofocusing and hydroxyapatite-HPLC. The lithocholate binding affinity of Y' protein (Kd < 0.35 microM) was higher than that of Y protein (Kd = 2 microM) and was comparable to that of Z protein (Kd = 0.2 microM). The binding affinity of Y protein was higher for bilirubin (Kd = 2.5 microM) than that for BSP (Kd = 200 microM). This was comparable to the binding affinity of bovine hepatic Y protein. These data indicate that Y' and Z proteins participate in the intracellular transport of bile acids from the brush border to the basolateral pole in enterocytes.
Takikawa
H
H
Department of Medicine, Teikyo University School of Medicine, Tokyo, Japan.
Arai
S
S
Yamanaka
M
M
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000378:metabolism; D017886:Durapatite; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006882:Hydroxyapatites; D007082:Ileum; D007413:Intestinal Mucosa / Q000378:metabolism*; D007700:Kinetics; D008095:Lithocholic Acid / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D011485:Protein Binding; D029701:Saccharomyces cerevisiae Proteins
D002352:Carrier Proteins; D005656:Fungal Proteins; D006882:Hydroxyapatites; C075720:PAU19 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D008095:Lithocholic Acid; D017886:Durapatite