Apical and basolateral parathyroid hormone receptors in rat renal cortical membranes.
Brush border (BBM) and basolateral membranes (BLM) of rat renal cortical cells separated by free flow electrophoresis revealed two distinct peaks of BBM-specific leucine aminopeptidase and Na+/K(+)-ATPase for BLM. PTH/PTH-related protein (PTHrP) receptors were identified in BBM and BLM. Specific binding of 125 pM [125I]chicken [Tyr36]-PTHrP-(1-36)amide [chPTHrP-(1-36)] to individual fractions of membranes separated by free flow electrophoresis overlapped with the leucine aminopeptidase and Na+/K(+)-ATPase profiles. Binding to pooled BBM was 53 +/- 5% (mean +/- SEM) of that to BLM (P < 0.01). In BBM and BLM, half-maximal inhibition of binding was obtained with 0.4-0.9 nM chPTHrP-(1-36) and 0.2-0.6 nM rat PTH-(1-34). Guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S; 100 microM) lowered chPTHrP-(1-36) binding to 50% of control levels, and half-maximal inhibition of binding was obtained with 480 and 8 nM GTP gamma S in BBM and BLM, respectively. Cross-linking of the PTH/PTHrP receptors with [125I]chPTHrP-(1-36) modified with N-hydroxysuccinimidyl-4-azidobenzoate revealed indistinguishable doublets of 83 and 73 kilodaltons in both BBM and BLM. Adenylyl cyclase was stimulated 6- and 10-fold by chPTHrP-(1-36) and GTP gamma S, respectively, in BLM and 1.3- and 1.9-fold in BBM. In conclusion, PTH receptors were recognized in both the basolateral and brush border membranes. Different receptor coupling to G-proteins and minimal cAMP stimulation in BBM provide evidence for PTH/PTHrP receptor isotypes and/or different postreceptor activation in BBM and BLM.
Kaufmann
M
M
Department of Orthopedic Surgery, University Hospital, Zurich, Switzerland.
Muff
R
R
Stieger
B
B
Biber
J
J
Murer
H
H
Fischer
J A
JA
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001485:Basement Membrane / Q000737:chemistry; D019204:GTP-Binding Proteins / Q000032:analysis; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007672:Kidney Cortex / Q000737:chemistry* / Q000648:ultrastructure; D008297:Male; D008871:Microvilli / Q000737:chemistry; D044162:Parathyroid Hormone-Related Protein; D010446:Peptide Fragments / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D018016:Receptors, Parathyroid Hormone / Q000032:analysis*
D044162:Parathyroid Hormone-Related Protein; D010446:Peptide Fragments; D011506:Proteins; D018016:Receptors, Parathyroid Hormone; C079957:parathyroid hormone-related peptide (1-36); D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases