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The proton translocating ATPase responsible for urinary acidification.

S Gluck, S Kelly, Q Al-Awqati

The Journal of Biological Chemistry

|August 25, 1982

Summary

The turtle urinary bladder uses a proton pump in its luminal membrane for acid secretion. Researchers identified an electrogenic proton-translocating ATPase responsible for this urinary acidification process.

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Area of Science:

Cell Biology
Physiology
Biochemistry

Background:

The turtle urinary bladder secretes acid (H+) to regulate urine pH.
This process is driven by a proton pump situated in the luminal membrane of bladder cells.

Purpose of the Study:

To identify and characterize the specific enzyme responsible for proton (H+) secretion in the turtle urinary bladder.
To confirm the localization of this proton pump within the cell.

Main Methods:

Isolation of a microsomal fraction from turtle urinary bladder cells.
Assay of proton translocating ATPase activity.
Inhibition studies using dicyclohexylcarbodiimide, oligomycin, and vanadate.
Sucrose density gradient centrifugation with labeling of luminal membranes (concanavalin A) and lysosomal markers.

Main Results:

An electrogenic proton translocating ATPase was found in the microsomal fraction.
This ATPase was inhibited by dicyclohexylcarbodiimide but not by oligomycin or vanadate.
The ATPase co-migrated with luminal membranes and was distinct from a lysosomal marker.

Conclusions:

The identified electrogenic proton translocating ATPase is the H+ ATPase responsible for urinary acidification in the turtle bladder.
This enzyme is localized to the luminal membrane, consistent with its role in H+ secretion.

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