Characterization of the ileal Na+/bile salt co-transporter in brush border membrane vesicles and functional expression in Xenopus laevis oocytes.
The Na+/bile salt co-transporter of the pig ileal brush border membrane has been expressed in Xenopus laevis oocytes. Injection of pig ileal poly (A)+ RNA into oocytes resulted in the functional expression of an Na(+)-gradient-stimulated taurocholate uptake within 2-5 days. The expressed Na(+)-dependent taurocholate uptake exhibited saturation kinetics (apparent Km of 48 microM), and displayed similar competitive substrate inhibition by taurodeoxycholate as the native brush border Na+/bile salt co-transporter studied in pig ileal brush border membrane vesicles. Interestingly, injection of pig proximal and mid intestinal poly (A)+ RNA into oocytes also resulted in the expression of the Na+/bile salt co-transporter, though the Na(+)-dependent transport of bile salts does not occur in brush border membrane vesicles (BBMV) isolated from pig proximal and mid intestine. This suggests that the mRNA coding for the co-transporter is present in the enterocytes lining the whole length of the small intestine, but that the function is only expressed in the brush border of the distal small intestine. The transport of D-glucose into BBMV, and the transport of methyl-alpha-D-glucopyranoside (a non-metabolizable hexose derivative) into oocytes were used throughout the study as methods of confirming the integrity of vesicles and oocytes.
Mullins
J G
JG
Department of Biochemistry, University of Wales, Aberystwyth, Dyfed, U.K.
Beechey
R B
RB
Gould
G W
GW
Campbell
F C
FC
Shirazi-Beechey
S P
SP
D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism; D001667:Binding, Competitive; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D006913:Hydroxysteroid Dehydrogenases; D007082:Ileum / Q000378:metabolism* / Q000648:ultrastructure; D007422:Intestines / Q000737:chemistry; D008081:Liposomes / Q000378:metabolism; D008562:Membrane Glycoproteins; D008871:Microvilli / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D012964:Sodium / Q000494:pharmacology*; D013552:Swine; D013656:Taurocholic Acid / Q000378:metabolism; D014162:Transfection; D014982:Xenopus laevis; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
D001647:Bile Acids and Salts; D002352:Carrier Proteins; D005947:Glucose; D006913:Hydroxysteroid Dehydrogenases; D008081:Liposomes; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D012964:Sodium; D013656:Taurocholic Acid; C037643:bile acid binding proteins; C435496:AKR1C2 protein, human; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)