Kidney brush-border membranes contain two sodium-dependent glucose transporters, one with low and one with high affinity for phlorizin, the specific inhibitor of these transporters. Using Scatchard analysis of phlorizin binding and Western blotting with specific antibodies against these transporters, we demonstrate in this study that although both transporters were proteolysed by papain treatment, only the high-affinity phlorizin-binding sites were decreased. Papain treatment followed by cross-linking with homobifunctional disuccinimidyl tartarate restored only the structure of the low-affinity phlorizin-binding protein (approx. molecular mass 70 kDa) without modifying the phlorizin-binding sites. When disuccinimidyl tartarate was replaced with dithiobis(succinimidyl acetate), another homobifunctional cross-linker with a higher spacer arm, the low- and high-affinity sites were both restored, with reappearance of two phlorizin-binding proteins with approx. molecular masses of 70 and 120 kDa. We conclude that high-affinity phlorizin-binding sites depend on the presence of the heterodimeric 120 kDa protein.
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Research Article|
March 01 1998
Effect of cross-linkers on the structure and function of pig-renal sodium–glucose cotransporters after papain treatment Available to Purchase
Jean GIUDICELLI;
Jean GIUDICELLI
1Laboratoire de Biochimie, Faculté de Médecine, Avenue de Valombrose, F-06107 Nice Cedex 2, France
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Marie-France BERTRAND;
Marie-France BERTRAND
1Laboratoire de Biochimie, Faculté de Médecine, Avenue de Valombrose, F-06107 Nice Cedex 2, France
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Stephane BILSKI;
Stephane BILSKI
1Laboratoire de Biochimie, Faculté de Médecine, Avenue de Valombrose, F-06107 Nice Cedex 2, France
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T. Than TRAN;
T. Than TRAN
1Laboratoire de Biochimie, Faculté de Médecine, Avenue de Valombrose, F-06107 Nice Cedex 2, France
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Jean-Claude POIREE
Jean-Claude POIREE
1
1Laboratoire de Biochimie, Faculté de Médecine, Avenue de Valombrose, F-06107 Nice Cedex 2, France
1To whom correspondence should be addressed.
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Publisher: Portland Press Ltd
Received:
July 21 1997
Revision Received:
October 20 1997
Accepted:
November 03 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 330 (2): 733–736.
Article history
Received:
July 21 1997
Revision Received:
October 20 1997
Accepted:
November 03 1997
Citation
Jean GIUDICELLI, Marie-France BERTRAND, Stephane BILSKI, T. Than TRAN, Jean-Claude POIREE; Effect of cross-linkers on the structure and function of pig-renal sodium–glucose cotransporters after papain treatment. Biochem J 1 March 1998; 330 (2): 733–736. doi: https://doi.org/10.1042/bj3300733
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