Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Phorbol ester-stimulated phosphorylation of basolateral membranes from canine kidney

Journal Article · · Am. J. Physiol.; (United States)
OSTI ID:5230579
; ; ;

To determine whether protein kinase C is present in the basolateral membrane of the renal proximal tubular cell, we performed experiments to ascertain whether specific binding of (/sup 3/H)phorbol 12,13-dibutyrate could be demonstrated in basolateral membranes isolated from canine kidney. Specific binding was demonstrable that was half maximal at between 10(-7) and 10(-8) M phorbol 12,13-dibutyrate. Binding was inhibited by 12-O-tetradecanoylphorbol-13-acetate (TPA) and other tumor-promoting phorbol esters, but not by inactive phorbol esters, including 4 alpha-phorbol. Incubation of basolateral membranes with TPA and phorbol 12,13-dibutyrate, but not with 4 alpha-phorbol, in the presence of submicromolar concentrations of free calcium, enhanced phosphorylation of several proteins demonstrable in autoradiograms of sodium dodecyl sulfate-polyacrylamide gels originating from membranes subsequently exposed to (gamma-32P)ATP for 30 s. Dephosphorylation of (/sup 32/P)phosphoproteins was observed in gels from membranes incubated with (gamma-32P)ATP over time. TPA-stimulated phosphorylation of one protein band with Mr 135,000 was quantitated and was found to increase as a function of (TPA). Half-maximal TPA-stimulated phosphorylation of this protein band occurred at slightly less than 10(-9) M TPA. Our findings are consistent with a role for protein kinase C-effected phosphorylation of basolateral membrane proteins in the mediation or modulation of hormonal actions in the proximal tubular cell.

Research Organization:
Washington Univ. School of Medicine, St. Louis, MO
OSTI ID:
5230579
Journal Information:
Am. J. Physiol.; (United States), Journal Name: Am. J. Physiol.; (United States); ISSN AJPHA
Country of Publication:
United States
Language:
English

Similar Records

Phorbol esters inhibit ammoniagenesis and gluconeogenesis in proximal tubular segments
Journal Article · Mon Jun 01 00:00:00 EDT 1987 · Am. J. Physiol.: Heart Circ. Physiol.; (United States) · OSTI ID:6201295
Binding of IGF I and IGF I-stimulated phosphorylation in canine renal basolateral membranes
Journal Article · Tue Jul 01 00:00:00 EDT 1986 · Am. J. Physiol.; (United States) · OSTI ID:5092828
Tumor-promoting phorbol esters effect alkalinization of canine renal proximal tubular cells
Conference · Fri Feb 28 23:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) · OSTI ID:5388302

Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BODY
CARCINOGENS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL BONDS
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
DOGS
ENZYME ACTIVITY
ENZYMES
ESTERS
IN VITRO
ISOTOPE APPLICATIONS
ISOTOPES
KIDNEYS
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPIDS
MAMMALS
MEMBRANES
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHORBOL ESTERS
PHOSPHOLIPIDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
RADIOISOTOPES
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES