Localization of phosphoinositide-specific phospholipase C-alpha in porcine kidney.
The presence of multiple forms of phosphoinositide-specific phospholipase C (PLC), an important enzyme in the cell signal transduction, suggests that specialized functions of tissues and cells may require different modes of PLC regulation. In the present study, we have purified a 54-kDa heparin-binding protein from a 4 M guanidine hydrochloride extract of porcine kidney, and identified it as one of isoenzymes of PLC on the basis of its partial amino acid sequence. Among 194 determined sequences of the porcine protein, 186 residues were identical with those deduced from nucleotide sequence of the cDNA encoding rat PLC-alpha. The subcellular distribution of porcine renal PLC-alpha was examined by Western blotting by using a specific antibody against the purified protein. Quantitation of the Western blots revealed that 70% of PLC-alpha was membrane-associated. Immunohistochemical studies showed a specific localization of PLC-alpha in epithelial cells of distal tubules and collecting ducts of normal porcine kidney, but not in other cells composing the nephron. Moreover, the highest expression of PLC-alpha was observed in apical membranes in these epithelial cells. Thus, this form of PLC is considered to have a specific role in the signal transduction process related to regional renal tubular functions.
Wakui
H
H
Third Department of Internal Medicine, Akita University School of Medicine, Japan.
Komatsuda
A
A
Ishino
T
T
Imai
H
H
Kobayashi
R
R
Nakamoto
Y
Y
Miura
A B
AB
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis; D006493:Heparin / Q000378:metabolism; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000302:isolation & purification*; D007668:Kidney / Q000201:enzymology*; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism*; D013552:Swine; D010738:Type C Phospholipases / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*
D002352:Carrier Proteins; D007527:Isoenzymes; D010716:Phosphatidylinositols; D006493:Heparin; D010738:Type C Phospholipases