Interactions of (/sup 14/C)phosphonoformic acid with renal cortical brush-border membranes. Relationship to the Na+-phosphate co-transporter
Since phosphonoformic acid (PFA) acts as a specific competitive inhibitor of Na+-Pi co-transport across renal brush-border membrane (BBM), we employed the (/sup 14/C)PFA as a probe to determine the mechanism of its interaction with rat renal BBM. The binding of (/sup 14/C)PFA to BBM vesicles (BBMV), with Na+ present in extravesicular medium (Na+o), was time- and temperature-dependent. The replacement of Na+o with other monovalent cations reduced the PFA binding by -80%. Cl- was the most effective accompanying monovalent anion as NaCl for maximum PFA binding. The Na+o increased the apparent affinity of BBMV for (/sup 14/C)PFA binding, but it did not change the maximum binding capacity. The maximum (/sup 14/C)PFA binding was achieved at Na+o approximately equal to 50 mM. The extent of Na+-dependent (/sup 14/C)PFA binding correlated with percent inhibition by an equimolar dose of PFA of the dependent BBMV uptake of 32Pi. Intravesicular Na+ (Na+i) decreased (/sup 14/C)PFA binding, on BBMV, and this inhibition by Na+i was dependent on the presence of Na+o. The increase in Na+i, at constant (Na+)o, decreased the Vmax, but not the Km, for (/sup 14/C)PFA binding on BBMV. Bound (/sup 14/C)PFA was displaced from BBMV by phosphonocarboxylic acids proportionally to their ability to inhibit gradient-dependent Pi transport, whereas other monophosphonates, diphosphonates, L-proline, or D-glucose did not influence the (/sup 14/C)PFA binding. The Na+-dependent binding of (/sup 14/C)PFA and of (/sup 3/H)phlorizin by BBMV was 10 times higher than binding of these ligands to renal basolateral membranes and to mitochondria. (/sup 14/C)PFA probably binds onto the same locus on the luminal surface of BBM, where Pi and Na+ form a ternary complex with the Na+-Pi co-transporter.
- Research Organization:
- Mayo Clinic, Rochester, MN
- OSTI ID:
- 6075518
- Journal Information:
- J. Biol. Chem.; (United States), Vol. 262:17
- Country of Publication:
- United States
- Language:
- English
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550201* - Biochemistry- Tracer Techniques