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  Single Molecule Recognition of Protein Binding Epitopes in Brush Border Membranes by Force Microscopy

Wielert-Badt, S., Hinterdorfer, P., Gruber, H. J., Lin, J.-T., Badt, D., Wimmer, B., et al. (2002). Single Molecule Recognition of Protein Binding Epitopes in Brush Border Membranes by Force Microscopy. Biophysical Journal, 82(5): 1, pp. 2767-2774. Retrieved from http://www.biophysj.org/cgi/content/abstract/82/5/2767.

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Genre: Journal Article
Alternative Title : Biophys. J.

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 Creators:
Wielert-Badt, Susanne1, Author
Hinterdorfer, Peter, Author
Gruber, Hermann J., Author
Lin, Jiann-Trzuo2, Author              
Badt, Dirk, Author
Wimmer, Barbara1, Author
Schindler, Hansgeorg, Author
Kinne, Rolf K. H.2, Author              
Affiliations:
1Max Planck Institute of Molecular Physiology, Max Planck Society, ou_1753286              
2Sonstige Wissenschaftliche Organisationseinheiten, Max Planck Institute of Molecular Physiology, Max Planck Society, ou_1753294              

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 Abstract: Sidedness and accessibility of protein epitopes in intact brush border membrane vesicles were analyzed by detecting single molecule interaction forces using molecular recognition force microscopy in aqueous physiological solutions. Frequent antibody-antigen recognition events were observed with a force microscopy tip carrying an antibody directed against the periplasmically located gamma-glutamyltrans- peptidase, suggesting a right side out orientation of the vesicles. Phlorizin attached to the tips bound to NA(+)/D-glucose cotransporter molecules present in the vesicles. The recognition was sodium dependent and inhibited by free phlorizin and D-glucose, and revealed an apparent K-D of 0.2 muM. Binding events were also observed with an antibody directed against the epitope aa603-aa630 close to the C terminus of the transporter. In the presence of phlorizin the probability of antibody binding was reduced but the most probable unbinding force f(u) = 100 pN remained unchanged. In the presence of D-glucose and sodium, however, both the binding probability and the most probable binding force (f(u) = 50 pN) were lower than in its absence. These studies demonstrate that molecular recognition force microscopy is a versatile tool to probe orientation and conformational changes of epitopes of membrane components during binding and traps-membrane transpor

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Language(s): eng - English
 Dates: 2002-05
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Type: Peer
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Title: Biophysical Journal
  Alternative Title : Biophys. J.
Source Genre: Journal
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Pages: - Volume / Issue: 82 (5) Sequence Number: 1 Start / End Page: 2767 - 2774 Identifier: -