Endogenous glycosylphosphatidylinositol-specific phospholipase C releases renal dipeptidase from kidney proximal tubules in vitro.
Spontaneous enzymic release of renal dipeptidase (RDPase; EC 3.4.13.19), a glycosylphosphatidylinositol (GPI)-linked ectoenzyme, was observed in vitro during incubation of porcine proximal tubules at 37 degrees C. Triton X-114 phase separation of the released RDPase showed that the majority of the enzyme activity partitioned into the aqueous phase, indicating its hydrophilic nature. Immunoblot analyses using an antibody against the cross-reacting determinant (CRD) inositol 1,2-cyclic monophosphate, the epitope formed by phospholipase C (PLC) cleavage of the GPI anchor on a protein, detected the released RDPase. Reprobing the immunoblot with an anti-RDPase serum showed the RDPase band co-migrating with the CRD band. The release of RDPase from the proximal tubules was a Ca(2+)-dependent process and had a pH optimum of 9.0. These results indicate that RDPase is released from the proximal tubules by the action of a distinct endogenous GPI-specific PLC.
Park
S W
SW
College of Pharmacy, Chonnam National University, Kwangju 500-757, Korea.
Choi
K
K
Kim
I C
IC
Lee
H H
HH
Hooper
N M
NM
Park
H S
HS
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004150:Dipeptidases / Q000737:chemistry / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000201:enzymology*; D017830:Octoxynol; D011092:Polyethylene Glycols; D012995:Solubility; D013552:Swine; D010738:Type C Phospholipases / Q000378:metabolism*
D017261:Glycosylphosphatidylinositols; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D010738:Type C Phospholipases; D004150:Dipeptidases; C038937:dipeptidase; D002118:Calcium