Biosynthesis of the cloned intestinal Na+/glucose cotransporter

Functional Regulation of Sugar Assimilation by N-Glycan-specific Interaction of Pancreatic α-Amylase with Glycoproteins of Duodenal Brush Border Membrane

“…A single N-glycosylation site is utilized at Asn-248 (51). Previously, it was reported that the glycosylation of SGLT1 is not necessary for transport of Glc because deglycosylation of BBMV (52) and deletion of potential N-glycosylation sites by mutation did not reduce the Glc transport activity of SGLT1 (38,51). In this study, unexpectedly, Glc uptake by SGLT1 was almost completely inhibited by PPA, which was attenuated only by a mannose-specific lectin, GNA (Fig.…”

The Effect of Massive Small Bowel Resection and Oral Epidermal Growth Factor Therapy on SGLT-1 Distribution in Rabbit Distal Remnant

“…Analysis of BBM sugar transporter levels from unmanipulated control and transected control groups did not reveal any significant differences, therefore control data were pooled for ease of presentation. Probing with the polyclonal antibody against SGLT-1 revealed one immunoreactive band at roughly 70 -75 kD, which is in agreement with previous reports for rabbit 13,14). As shown in Figure 1, BBM from remnant tissues contained less SGLT-1 than compared with control BBM (resected, 0.347 Ϯ 0.041, n ϭ 10, versus pooled controls, 0.487 Ϯ 0.041 arbitrary units trace OD, n ϭ 10, p Ͻ 0.01).…”

“…Poly-A ϩ RNA (mRNA) was purified using the polyASpin mRNA Isolation Kit (polyASpin Isolation Kit, 1560; New England Biolabs, Mississauga, Ontario, Canada). An oligo-dT primed (homo-oligomeric DNA pd (T) [12][13][14][15][16][17][18] , 2778503; Pharmacia Biotech, Baie D'Urfe, Quebec, Canada) first-strand cDNA was prepared using Superscript II reverse transcriptase (SuperScript II RNase H-reverse transcriptase, 18064022; GIBCO Life Technologies) and 2 g of poly-A ϩ 20 RNA from a control rabbit as template. A full-length SGLT-1 cDNA was prepared by PCR using primers based on the published rabbit SGLT-1 mRNA sequence obtained from Genbank (accession number M84020), and was subsequently cloned into pBluescript SK (Stratagene, La Jolla, CA, U.S.A.).…”

Identification and Functional Characterization of a Na+-independent Neutral Amino Acid Transporter with Broad Substrate Selectivity

“…The transmembrane regions of proteins were predicted on the amino acid sequence based on the SOSUI algorithm (28). In vitro translation of cRNA was performed as described elsewhere (18,29,30).…”