gamma-L-glutamyl-L-DOPA inhibits Na(+)-phosphate cotransport across renal brush border membranes and increases renal excretion of phosphate.
BACKGROUND
For treatment of phosphate (Pi) overload in various pathophysiological states, an agent that selectively increases renal Pi excretion would be of major value. Previously, we have shown that dopamine (DA) inhibits Na(+)-Pi cotransport in renal epithelia. However, the administration of DA or its immediate precursor L-DOPA increases DA in multiple tissues. Synthetic dipeptide gamma-L-glutamyl-L-DOPA (gludopa) can serve as an inactive precursor (pro-pro-drug) of DA. This study tested the hypothesis that, because of the unique colocalization of gamma-glutamyltransferase (gamma-GT), aromatic amino acid decarboxylase, Na(+)-Pi cotransporter, and Na(+)-L-DOPA cotransporter in brush border membrane (BBM) of proximal tubular cells, gludopa may elicit phosphaturia by action of DA generated within the kidney.


METHODS
Thyroparathyrectomized rats were given placebo, or gludopa, or gludopa + gamma-GT inhibitor acivicin. Urinary excretion of Pi, Ca2+, Na+, K+, DA, cAMP, and cGMP was determined, and Na(+)-Pi cotransport was measured in BBM prepared from kidneys of rats at the end of the experiment.


RESULTS
The administration of gludopa resulted in: (a) an inhibition of Na(+)-Pi cotransport, but not cotransport of Na(+)-proline and Na(+)-alanine in BBM; (b) an increase (+300%) of fractional excretion (FE) of Pi and a drop (-35%) of plasma Pi, whereas the plasma levels and FEs of Ca2+, Na+, and K+ were unchanged; (c) an increase in urinary excretion of cAMP. but not cGMP; (d) a 1000-fold increase of urinary excretion of DA, without a change in excretion of norepinephrine; and (e) an incubation of gludopa with BBM in vitro, which caused a release of L-DOPA, and the in vivo administration of acivicin, which blocked actions of gludopa to inhibit Na(+)-Pi cotransport and to increase urinary excretions of Pi and DA.


CONCLUSIONS
We conclude that colocalization of enzymes of biotransformation, BBM transporters, and the autocrine/paracrine DA system in cells of proximal tubules constitutes a cellular basis for the potent and specific phosphaturic action of gludopa.
de Toledo
F G
FG
Department of Physiology and Biophysics, Mayo Clinic and Foundation, Mayo Medical School, Rochester, Minnesota, USA.
Thompson
M A
MA
Bolliger
C
C
Tyce
G M
GM
Dousa
T P
TP
D000409:Alanine / Q000493:pharmacokinetics; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002118:Calcium / Q000097:blood / Q000652:urine; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D000242:Cyclic AMP / Q000652:urine; D006152:Cyclic GMP / Q000652:urine; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004298:Dopamine / Q000652:urine; D007687:Kidney Tubules, Proximal / Q000737:chemistry / Q000201:enzymology*; D007980:Levodopa / Q000097:blood / Q000652:urine; D008297:Male; D008871:Microvilli / Q000737:chemistry / Q000201:enzymology; D009638:Norepinephrine / Q000652:urine; D016105:Parathyroidectomy; D010710:Phosphates / Q000097:blood / Q000652:urine*; D011188:Potassium / Q000097:blood / Q000652:urine; D011392:Proline / Q000493:pharmacokinetics; D051381:Rats; D012964:Sodium / Q000097:blood / Q000652:urine; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D014316:Tritium; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D002352:Carrier Proteins; D010710:Phosphates; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D014316:Tritium; D007980:Levodopa; C017342:gamma-glutamyl DOPA; D004295:Dihydroxyphenylalanine; D011392:Proline; D012964:Sodium; D000242:Cyclic AMP; D005723:gamma-Glutamyltransferase; D006152:Cyclic GMP; D000409:Alanine; D011188:Potassium; D002118:Calcium; D004298:Dopamine; D009638:Norepinephrine