DOI: 10.1073/pnas.87.4.1456. Intestinal brush border membrane Na+/glucose cotransporter functions in situ as a homotetramer.

Intestinal brush border membrane Na+/glucose cotransporter functions in situ as a homotetramer.

10.1073/pnas.87.4.1456

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The functional unit molecular size of the intestinal brush border membrane-bound Na+/glucose cotransporter was determined by radiation inactivation. Purified brush border membrane vesicles preserved in cryoprotectant buffer were irradiated (-135 degrees C) with high-energy electrons from a 13-MeV (1 eV = 1.602 x 10(-19) J) linear accelerator at doses from 0 to 70 Mrad (1 rad = 0.01 Gy). After each dose, the cotransporter was investigated with respect to (i) Na(+)-dependent transport activity and (ii) immunologic blot analysis with antibodies against the cloned rabbit intestinal cotransporter. Increasing radiation decreased the maximal Na(+)-dependent cotransporter activity Jmax without affecting apparent Km. The size of the transporting functional unit was 290 +/- 5 kDa. Immunologic blot analysis of brush border membranes gave a single band of Mr 70,000, which decreased in intensity with increased radiation dose and gave a target size of 66 +/- 11 kDa. We conclude that activity of the intestinal Na+/glucose cotransporter in situ in the brush border membrane requires the simultaneous presence of four intact, independent, identical subunits arranged as a homotetramer.

Bibliography

Stevens, B. R., Fernandez, A., Hirayama, B., Wright, E. M., & Kempner, E. S. (1990). Intestinal brush border membrane Na+/glucose cotransporter functions in situ as a homotetramer. Proceedings of the National Academy of Sciences, 87(4), 1456â1460.

Authors 5

B R Stevens (first)
A Fernandez (additional)
B Hirayama (additional)
E M Wright (additional)
E S Kempner (additional)

References 0 Referenced 50

None

Dates

Type	When
Created	19 years, 3 months ago (May 31, 2006, 7:33 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 1:05 p.m.)
Indexed	1 month, 2 weeks ago (July 14, 2025, 11:33 p.m.)
Issued	35 years, 6 months ago (Feb. 1, 1990)
Published	35 years, 6 months ago (Feb. 1, 1990)
Published Online	35 years, 6 months ago (Feb. 1, 1990)
Published Print	35 years, 6 months ago (Feb. 1, 1990)

Funders 0

None

BibTeX

@article{Stevens_1990, title={Intestinal brush border membrane Na+/glucose cotransporter functions in situ as a homotetramer.}, volume={87}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.87.4.1456}, DOI={10.1073/pnas.87.4.1456}, number={4}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Stevens, B R and Fernandez, A and Hirayama, B and Wright, E M and Kempner, E S}, year={1990}, month=feb, pages={1456–1460} }

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