Association of protein kinase C(lambda) with adducin in 3T3-L1 adipocytes.
There is evidence that the atypical protein kinases C (PKC(lambda), PKC(zeta)) participate in signaling from the insulin receptor to cause the translocation of glucose transporters from an intracellular location to the plasma membrane in adipocytes. In order to search for downstream effectors of these PKCs, we identified the proteins that were immunoprecipitated by an antibody against PKC(lambda/zeta) from lysates of 3T3-L1 adipocytes through peptide sequencing by mass spectrometry. The data show that PKC(lambda) is the major atypical PKC in these cells. Moreover, an oligomeric complex consisting of alpha- and gamma-adducin, which are cytoskeletal proteins, coimmunoprecipitated with PKC(lambda). Association of the adducins with PKC(lambda) was further indicated by the finding that the adducins coimmunoprecipitated proportionally with PKC(lambda) in repeated rounds of immunoprecipitation. Such an association is consistent with literature reports that the adducins contain a single major site for PKC phosphorylation in their carboxy termini. Using antibody against the phospho form of this site for immunoblotting, we found that insulin caused little or no increase in the phosphorylation of this site on the adducins in a whole cell lysate or on the small portion of the adducins that coimmunoprecipitated with PKC(lambda). PKC(lambda) and the adducins were located in both the cytosol and subcellular membranous fractions. The binding of PKC(lambda) to adducin may function to localize PKC(lambda) in 3T3-L1 adipocytes.
Laustsen
P G
PG
Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755, USA.
Lane
W S
WS
Bennett
V
V
Lienhard
G E
GE
D016475:3T3 Cells; D017667:Adipocytes / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007328:Insulin / Q000494:pharmacology; D007527:Isoenzymes; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000032:analysis; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism; D000099291:Adducins; D000099293:Protein Kinase C-lambda
D002148:Calmodulin-Binding Proteins; D007328:Insulin; D007527:Isoenzymes; D011493:Protein Kinase C; D011506:Proteins; D000099291:Adducins; D000099293:Protein Kinase C-lambda