English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Proton ATPase in rat renal cortical endocytotic vesicles

MPS-Authors
/persons/resource/persons256395

Sabolic,  Ivan
Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society;

/persons/resource/persons251310

Burckhardt,  Gerhard
Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society;

External Resource
No external resources are shared
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Sabolic, I., & Burckhardt, G. (1988). Proton ATPase in rat renal cortical endocytotic vesicles. Biochimica et Biophysica Acta-Biomembranes, 937(2), 398-410. doi:10.1016/0005-2736(88)90262-3.


Cite as: http://hdl.handle.net/21.11116/0000-0007-DE19-4
Abstract
To relate ATPase activity to the ATP-driven H+-pump in rat renal endocytotic vesicles we applied an in vitro coupled optical test and a Pi-liberation assay. Endocytotic vesicles contain an ouabain-, vanadate- and oligomycin-insensitive ATPase. The ionophores for K+ and H+, valinomycin and carbonylcyanide p-chloro-methoxyphenylhydrazone (CCCP), respectively, stimulated ATPase activity, indicating its relation to the electrogenic H+-pump. This conclusion is supported by a similar distribution on a Percoll gradient of ATP-driven H+ uptake into endosomes and ionophore-stimulated ATPase activity. Coupled optical and Pi-liberation assays were then used to characterize the H+-ATPase with respect to the requirement for pH, nucleotides, anions, and mono- and divalent cations. The H+-ATPase activity was decreased by widely used blockers: N-ethylmaleimide (NEM), dicyclohexylcarbodiimide (DCCD) and diethylstilbestrol (DES). Different sensitivities to these blockers proved that alkaline phosphatase and H+-ATPase are separate entities. To investigate whether the NEM-, DCCD- and DES-sensitive ATPase activity is confined to intact endocytotic vesicles, cellular membranes from rat kidney cortex were separated on a Percoll density gradient. Surprisingly, endocytotic vesicles contain only a small fraction of the total NEM-, DCCD- and DES-sensitive ATPase activity. The majority of the blocker-sensitive ATPases belongs to membranes of as yet undefined cellular origin.