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  ATP-driven proton transport in vesicles from the kidney cortex

Sabolić, I., & Burckhardt, G. (1990). ATP-driven proton transport in vesicles from the kidney cortex. In S. Fleischer, & B. Fleischer (Eds.), Methods in Enyzmology (pp. 505-520). Academic Press. doi:10.1016/0076-6879(90)91033-3.

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 Creators:
Sabolić, Ivan1, Author           
Burckhardt, Gerhard1, Author           
Affiliations:
1Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068297              

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 Abstract: Publisher Summary:
This chapter describes an improved preparation of endocytotic vesicles from rat renal cortex. This method yields vesicles that are more enriched in proton pump activity. The preparation of endocytotic vesicles takes about 6 hr and does not require an ultracentrifuge. Therefore, endocytotic vesicles from kidney can serve as an excellent model to study the characteristics of the vacuolar proton pump (H+-ATPase). This chapter describes the way of tissue homogenization is of critical importance for the success of the preparation. Although the proton pump (H+- adenosine tri-phosphate (ATP)-ase) in isolated endocytotic vesicles is strongly inhibited by N,N'-dicyclohexylcarbodiimide (DCCD), N-Ethylmaleimide (NEM), and diethylstilbestrol (DES), neither of these inhibitors can be used as a specific marker for the pump. Experiments showing the distribution of inhibitor-sensitive ATPases indicate that much higher activities of DCCD-, NEM-, and DES-sensitive ATPases are present in fractions of the Percoll gradient that do not contain endocytotic vesicles with an ATP-driven proton pump.

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Language(s): eng - English
 Dates: 2004-01-071990
 Publication Status: Issued
 Pages: 15
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/0076-6879(90)91033-3
PMID: 1963653
 Degree: -

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Title: Methods in Enyzmology
  Subtitle : Biomembranes Part V: Cellular and Subcellular Transport: Epithelial Cells
Source Genre: Series
 Creator(s):
Fleischer, Sidney, Editor
Fleischer, Becca , Editor
Affiliations:
-
Publ. Info: Academic Press
Pages: 939 Volume / Issue: 191 Sequence Number: - Start / End Page: 505 - 520 Identifier: ISBN: 978-0-12-182092-3
ISSN: 0076-6879