Insulin controls subcellular localization and multisite phosphorylation of the phosphatidic acid phosphatase, lipin 1.
Brain, liver, kidney, heart, and skeletal muscle from fatty liver dystrophy (fld/fld) mice, which do not express lipin 1 (lipin), contained much less Mg(2+)-dependent phosphatidic acid phosphatase (PAP) activity than tissues from wild type mice. Lipin harboring the fld(2j) (Gly(84) --> Arg) mutation exhibited relatively little PAP activity. These results indicate that lipin is a major PAP in vivo and that the loss of PAP activity contributes to the fld phenotype. PAP activity was readily detected in immune complexes of lipin from 3T3-L1 adipocytes, where the protein was found both as a microsomal form and a soluble, more highly phosphorylated, form. Fifteen phosphorylation sites were identified by mass spectrometric analyses. Insulin increased the phosphorylation of multiple sites and promoted a gel shift that was due in part to phosphorylation of Ser(106). In contrast, epinephrine and oleic acid promoted dephosphorylation of lipin. The PAP-specific activity of lipin was not affected by the hormones or by dephosphorylation of lipin with protein phosphatase 1. However, the ratio of soluble to microsomal lipin was markedly increased in response to insulin and decreased in response to epinephrine and oleic acid. The results suggest that insulin and epinephrine control lipin primarily by changing localization rather than intrinsic PAP activity.
Harris
Thurl E
TE
Department of Pharmacology, University of Virginia, Charlottesville, Virginia 22908-0735, USA.
Huffman
Todd A
TA
Chi
An
A
Shabanowitz
Jeffrey
J
Hunt
Donald F
DF
Kumar
Anil
A
Lawrence
John C
JC
D041721:3T3-L1 Cells; D000818:Animals; D001120:Arginine / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D018014:Gene Transfer Techniques; D005998:Glycine / Q000737:chemistry; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins / Q000737:chemistry*; D019301:Oleic Acid / Q000737:chemistry; D010641:Phenotype; D010711:Phosphatidate Phosphatase; D010766:Phosphorylation; D011485:Protein Binding
D007328:Insulin; D009687:Nuclear Proteins; D019301:Oleic Acid; D001120:Arginine; C485999:LPIN1 protein, human; C418978:Lpin1 protein, mouse; D010711:Phosphatidate Phosphatase; D005998:Glycine