Control of endogenous phosphorylation of the major cAMP-dependent protein kinase substrate in adipocytes by insulin and beta-adrenergic stimulation.

In isolated, 32Pi-loaded, rat adipocytes, we have examined phosphorylation of the major cAMP-dependent protein kinase (A-kinase) substrate, a protein that appears to be associated with the lipid storage droplet and migrates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a 65-67-kDa doublet. In control cells, a strong phosphorylation signal is detected as the (+/- cAMP) A-kinase activity ratio ranges from approximately 0.1 to approximately 0.3-0.4 with increasing isoproterenol concentrations. By contrast, insulin-treated cells exhibiting A-kinase activity ratios over the range of 0.1-0.25 contain less 32P in the 65-67-kDa protein than control cells exhibiting identical A-kinase activity ratios. At higher activity ratios (greater than 0.3), this reduction in phosphorylation of the 65-67-kDa protein by insulin disappears. It is concluded that insulin stimulates a phosphatase activity that acts on the 65-67-kDa protein. Insulin actions aside, these studies reveal two interesting phenomena. 1) Whereas elevated, steady-state A-kinase activities are established rapidly (1-2 min) upon isoproterenol stimulation, phosphorylation of the 65-67-kDa substrate proceeds through a burst, followed by a decline to a steady-state level by 10-12 min. An "adaptation" mechanism, providing for a constant response to a constant stimulus, may underlie this lack of parallelism between the time course of phosphorylation and A-kinase activity. 2) Removal of [32Pi] orthophosphate immediately before isoproterenol stimulation leads to a rapid (t approximately 10 min) loss in labeling of the 65-67-kDa protein, whereas the phosphorylation state of other phosphoproteins are not changed. These data suggest that elevation of A-kinase activity leads to a rapid exchange of external Pi with an ATP pool that is used by A-kinase.

• Publication year

  1990

• Venue

  Journal of Biological Chemistry

• Publication date

  1990-11-05

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/S0021-9258(17)30578-1PMID 2172232
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Insulin-induced dephosphorylation of hormone-sensitive lipase. Correlation with lipolysis and cAMP-dependent protein kinase activity.

  1989cited by this paper
• Insulin stimulates dephosphorylation of phosphorylase in rat epitrochlearis muscles.

  1989cited by this paper
• Role of hormone-sensitive low Km cAMP phosphodiesterase in regulation of cAMP-dependent protein kinase and lipolysis in rat adipocytes.

  1989cited by this paper
• Differential regulation of S6 phosphorylation by insulin and epidermal growth factor in Swiss mouse 3T3 cells: insulin activation of type 1 phosphatase.

  1988cited by this paper
• Stimulation of protein phosphatase activity by insulin and growth factors in 3T3 cells.

  1988cited by this paper
• Activation of phosphofructokinase 2 by insulin in cultured hepatocytes without accompanying changes of effector levels or cAMP-stimulated protein kinase activity ratios.

  1988cited by this paper
• Insulin stimulates the dephosphorylation and activation of acetyl-CoA carboxylase.

  1988cited by this paper
• Phosphorylation of phosphoprotein phosphatase inhibitor-2 (I-2) in rat fat cells.

  1988cited by this paper
• Insulin action rapidly decreases multifunctional protein kinase activity in rat adipose tissue.

  1988cited by this paper
• A tandem chromatographic column method for assaying cAMP-dependent protein kinase and protein kinase C with synthetic peptide substrates.

  1988cited by this paper
• Studies on the specific activity of [gamma-32P]ATP in adipose and other tissue preparations incubated with medium containing [32P]phosphate.

  1986cited by this paper
• Early events in insulin actions.

  1986cited by this paper
• cAMP-dependent protein kinase and lipolysis in rat adipocytes. III. Multiple modes of insulin regulation of lipolysis and regulation of insulin responses by adenylate cyclase regulators.

  1985cited by this paper
• cAMP-dependent protein kinase and lipolysis in rat adipocytes. I. Cell preparation, manipulation, and predictability in behavior.

  1985cited by this paper
• cAMP-dependent protein kinase and lipolysis in rat adipocytes. II. Definition of steady-state relationship with lipolytic and antilipolytic modulators.

  1985cited by this paper
• Activation of glycogen synthase by insulin in rat adipocytes. Evidence of hormonal stimulation of multisite dephosphorylation by glucose transport-dependent and -independent pathways.

  1984cited by this paper
• Hormonal regulation of hormone-sensitive lipase in intact adipocytes: identification of phosphorylated sites and effects on the phosphorylation by lipolytic hormones and insulin.

  1984cited by this paper
• Determination of the cAMP-dependent protein kinase activity ratio in intact tissues.

  1983cited by this paper
• Hormonal regulation of protein dephosphorylation. Identification and hormonal regulation of protein phosphatase inhibitor-1 in rat adipose tissue.

  1983cited by this paper
• Measurement of hormone-stimulated protein phosphorylation in intact cells.

  1983cited by this paper
• Amplification and adaptation in regulatory and sensory systems.

  1982cited by this paper
• Hormone-sensitive lipase of rat adipose tissue. Purification and some properties.

  1981cited by this paper
• Hormone-sensitive lipase of rat adipose tissue: correlation of activity with a protein of molecular weight 84 000.

  1980cited by this paper
• Insulin stimulation of pyruvate dehydrogenase in an isolated plasma membrane-mitochondrial mixture occurs by activation of pyruvate dehydrogenase phosphatase.

  1980cited by this paper
• Subclasses of external adenosine receptors.

  1980cited by this paper
• Action of insulin and catecholamines on the phosphorylation of proteins associated with the cytosol, membranes, and "fat cake" of rat fat cells.

  1978cited by this paper
• Regulation by lipolytic and antillipolytic compounds of the phosphorylation of specific proteins in isolated intact fat cells.

  1976cited by this paper
• [43] Preparation of homogeneous cyclic AMP-dependent protein kinase(s) and its subunits from rabbit skeletal muscle

  1974cited by this paper
• Studies on the phosphorylation and activation of skeletal muscle phosphorylase and phosphorylase kinase in vivo.

  1970cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper
• The Metabolism of Isolated Fat Cells: I. Effects of Hormones on Glucose Metabolism and Lipolysis

  1964cited by this paper
• A Rapid Enzymatic Assay for Glycerol

  1962cited by this paper

• Perilipins: Current perspectives on the role in cardiovascular diseases

  2026cites this paper
• PLINs-mediated organelle interactions: a key of exercise-mediated improvement of skeletal muscle lipid metabolism disorders

  2025cites this paper
• Differential pathways to hepatic steatosis in fish: Divergent molecular mechanisms underlying high-carbohydrate versus high-lipid diet-induced lipid accumulation

  2025cites this paper
• Regulation of lipid droplets and cholesterol metabolism in adrenal cortical cells.

  2024cites this paper
• MIIP downregulation drives colorectal cancer progression through inducing peri-cancerous adipose tissue browning

  2024cites this paper
• Molecular mechanisms of perilipin protein function in lipid droplet metabolism

  2023cites this paper
• MIIP downregulation promotes colorectal cancer progression via inducing adjacent adipocytes browning

  2023cites this paper
• An Overview of Hormone-Sensitive Lipase (HSL)

  2022cites this paper
• The insulin centennial—100 years of milestones in biochemistry

  2021cites this paper
• The "discovery" of lipid droplets: A brief history of organelles hidden in plain sight.

  2020cites this paper
• Perilipin 5 S155 phosphorylation by PKA is required for the control of hepatic lipid metabolism and glycemic control

  2020cites this paper
• A phosphoprotein helps lipid droplets stay in shape

  2020cites this paper
• CB1 Receptor-Dependent and Independent Induction of Lipolysis in Primary Rat Adipocytes by the Inverse Agonist Rimonabant (SR141716A)

  2020cites this paper
• Nuclear Factor E2-Related Factor 2 Mediates Oxidative Stress-Induced Lipid Accumulation in Adipocytes by Increasing Adipogenesis and Decreasing Lipolysis

  2020cites this paper
• Cytological and functional characteristics of fascia adipocytes in rats: A unique population of adipocytes.

  2019cites this paper
• Sulfhydration of perilipin 1 is involved in the inhibitory effects of cystathionine gamma lyase/hydrogen sulfide on adipocyte lipolysis.

  2019cites this paper
• Porcine perilipin (PLIN) gene: Structure, polymorphism and association study in Large White pigs

  2018cites this paper
• Mechanisms of Insulin Action and Insulin Resistance.

  2018cites this paper
• Atorvastatin reduces lipid accumulation in the liver by activating protein kinase A-mediated phosphorylation of perilipin 5.

  2017cites this paper
• Methacrylated gelatin and mature adipocytes are promising components for adipose tissue engineering

  2016cites this paper
• New Analytical Tool for the Detection of Ractopamine Abuse in Goat Skeletal Muscle by Potential Gene Expression Biomarkers.

  2016cites this paper
• A Role for the Lipid Droplet Protein HIG2 in Promoting Lipid Deposition in Liver and Adipose Tissue: A Dissertation

  2016cites this paper
• Lipid Droplet Proteins and Hepatic Lipid Metabolism

  2016cites this paper
• Characteristics and functions of lipid droplets and associated proteins in enterocytes.

  2016cites this paper
• The Role of PDE3B Phosphorylation in the Inhibition of Lipolysis by Insulin

  2015cites this paper
• Unraveling the roles of PLIN5: linking cell biology to physiology.

  2015cites this paper
• Understanding the effects of mature adipocytes and endothelial cells on fatty acid metabolism and vascular tone in physiological fatty tissue for vascularized adipose tissue engineering

  2015cites this paper
• Cellular changes in lipid metabolism in response to altered energy balance and heat stress in lactating dairy cows

  2015cites this paper
• Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

  2015cites this paper
• Effects of acute heat stress on lipid metabolism of bovine primary adipocytes.

  2015cites this paper
• Untersuchungen zur Dynamik des Lipid-Metabolismus von Dictyostelium discoideum

  2014influential citation
• The Hepatitis C Virus Core Protein Inhibits Adipose Triglyceride Lipase (ATGL)-mediated Lipid Mobilization and Enhances the ATGL Interaction with Comparative Gene Identification 58 (CGI-58) and Lipid Droplets*

  2014influential citation
• The Interplay of Protein Kinase A and Perilipin 5 Regulates Cardiac Lipolysis*♦

  2014cites this paper
• Skeletal muscle PLIN3 and PLIN5 are serine phosphorylated at rest and following lipolysis during adrenergic or contractile stimulation

  2013cites this paper
• The role of skeletal muscle PLIN proteins at rest and following lipolytic stimulation

  2013cites this paper
• Skeletal muscle PLIN proteins, ATGL and CGI-58, interactions at rest and following stimulated contraction.

  2013cites this paper
• PPARγ coactivator-1α contributes to exercise-induced regulation of intramuscular lipid droplet programming in mice and humans

  2013cites this paper
• Increase or Decrease Hydrogen Sulfide Exert Opposite Lipolysis, but Reduce Global Insulin Resistance in High Fatty Diet Induced Obese Mice

  2013cites this paper
• Miglitol prevents diet-induced obesity by stimulating brown adipose tissue and energy expenditure independent of preventing the digestion of carbohydrates.

  2013cites this paper
• Lipid metabolism in skeletal muscle: generation of adaptive and maladaptive intracellular signals for cellular function.

  2012cites this paper
• Update on perilipin polymorphisms and obesity.

  2012cites this paper
• Lipolysis Response to Endoplasmic Reticulum Stress in Adipose Cells*

  2012influential citation
• Evidence for protein‐mediated fatty acid efflux by adipocytes

  2012cites this paper
• Identification of transcription factors and response elements driving expression of Plin5

  2012cites this paper
• Lipolysis response to endoplasmic reticulum stress in adipose cells.

  2012cites this paper
• The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis.

  2011cites this paper
• Apolipoprotein D, apolipoprotein R and ST6GalNAc4 genes respond to clenbuterol administration in pig adipose tissue

  2011cites this paper
• Polymerase I and Transcript Release Factor Regulates Lipolysis via a Phosphorylation-Dependent Mechanism

  2011cites this paper
• Central melanocortin stimulation increases phosphorylated perilipin A and hormone-sensitive lipase in adipose tissues.

  2010cites this paper
• Adoption of PERILIPIN as a unifying nomenclature for the mammalian PAT-family of intracellular lipid storage droplet proteins

  2010cites this paper
• Regulation of Energy Stores and Feeding by Neuronal and Peripheral CREB Activity in Drosophila

  2009cites this paper
• Regulation of adipose tissue lipolysis revisited

  2009cites this paper
• Symposium on ‘ Frontiers in adipose tissue biology ’ Regulation of adipose tissue lipolysis revisited

  2009cites this paper
• PAT proteins, an ancient family of lipid droplet proteins that regulate cellular lipid stores.

  2009cites this paper
• Perilipin is present in islets of Langerhans and protects against lipotoxicity when overexpressed in the beta-cell line INS-1.

  2009cites this paper
• Modulation of myocellular fat stores: lipid droplet dynamics in health and disease.

  2009cites this paper
• Resveratrol, a naturally occurring diphenolic compound, affects lipogenesis, lipolysis and the antilipolytic action of insulin in isolated rat adipocytes.

  2009cites this paper
• Deficiency of Adipose Differentiation-related Protein Impairs Foam Cell Formation and Deficiency of Adipose Differentiation-related Protein Impairs Foam Cell Formation and Protects against Atherosclerosis Materials and Methods Analysis of Atherosclerotic Lesions Electron Microscopy Analysis of Ather

  2008cites this paper
• Deficiency of Adipose Differentiation-Related Protein Impairs Foam Cell Formation and Protects Against Atherosclerosis

  2008cites this paper
• Mechanisms of metformin inhibiting lipolytic response to isoproterenol in primary rat adipocytes.

  2008cites this paper
• Perilipin polymorphism interacts with dietary carbohydrates to modulate anthropometric traits in hispanics of Caribbean origin.

  2008cites this paper
• Dietary effects on gene regulation and function in human adipose tissue in obesity

  2007influential citation
• Thematic review series: Adipocyte Biology. The perilipin family of structural lipid droplet proteins: stabilization of lipid droplets and control of lipolysis Published, JLR Papers in Press, September 18, 2007.

  2007cites this paper
• Differential gene expression profile in pig adipose tissue treated with/without clenbuterol

  2007cites this paper
• Hormonal Control of Reversible Translocation of Perilipin B to the Plasma Membrane in Primary Human Adipocytes*

  2006cites this paper
• Calyculin and okadaic acid promote perilipin phosphorylation and increase lipolysis in primary rat adipocytes.

  2006cites this paper
• Expanding role of caveolae in control of adipocyte metabolism : proteomics of caveolae

  2006cites this paper
• Insulin disrupts beta-adrenergic signalling to protein kinase A in adipocytes.

  2005cites this paper
• Activation of the Lipid Droplet Controls the Rate of Lipolysis of Triglycerides in the Insect Fat Body*

  2005cites this paper
• Role of PAT proteins in lipid metabolism.

  2005cites this paper
• Expression and localization of adipophilin and perilipin in human fetal membranes: association with lipid bodies and enzymes involved in prostaglandin synthesis.

  2005cites this paper
• Regulació nutricional i endocrina de la lipòlisi i la lipoproteïna lipasa en la truita irisada ("Oncorhynchus mykiss") i l'orada ("Sparus aurata")

  2005cites this paper
• Insulin disrupts β-adrenergic signalling to protein kinase A in adipocytes

  2005cites this paper
• Upregulated expression of stanniocalcin-1 during adipogenesis.

  2004cites this paper
• Appearance of smaller lipid bodies and protein kinase activation in the lipid body fraction are induced by an increase in the nitrogen source in the Mortierella fungus.

  2004cites this paper
• Triacylglycerol hydrolase: role in intracellular lipid metabolism

  2004cites this paper
• Adipose tissue expression of the lipid droplet-associating proteins S3-12 and perilipin is controlled by peroxisome proliferator-activated receptor-gamma.

  2004cites this paper
• Metabolism of lipids in human white adipocyte.

  2004cites this paper
• Insulin and Oleate Promote Translocation of Inosine‐5′ Monophosphate Dehydrogenase to Lipid Bodies

  2004cites this paper
• Identification of the lipid droplet-associated protein adipophilin as a novel liver x receptor target gene

  2004cites this paper
• Métabolisme lipidique dans le tissu adipeux blanc humain

  2004cites this paper
• Perilipin expression in human adipose tissues: effects of severe obesity, gender, and depot.

  2003cites this paper
• Analysis of lipolysis in adipocytes using a fluorescent fatty acid derivative.

  2003cites this paper
• Targeting of Big Stanniocalcin and Its Receptor to Lipid Storage Droplets of Ovarian Steroidogenic Cells*

  2003cites this paper
• The Central Domain Is Required to Target and Anchor Perilipin A to Lipid Droplets*

  2003cites this paper
• Lipase-selective Functional Domains of Perilipin A Differentially Regulate Constitutive and Protein Kinase A-stimulated Lipolysis*

  2003cites this paper
• Modulation of Hormone-sensitive Lipase and Protein Kinase A-mediated Lipolysis by Perilipin A in an Adenoviral Reconstituted System*

  2002cites this paper
• Tumor necrosis factor-alpha stimulates lipolysis in differentiated human adipocytes through activation of extracellular signal-related kinase and elevation of intracellular cAMP.

  2002cites this paper
• Stimulation of Lipolysis and Hormone-sensitive Lipase via the Extracellular Signal-regulated Kinase Pathway*

  2001cites this paper
• Serial changes in the myocardial β-adrenergic signalling system in two models of non-insulin dependent diabetes mellitus

  2001cites this paper
• The biogenesis and functions of lipid bodies in animals, plants and microorganisms.

  2001cites this paper
• Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation during the lactation cycle of the rat.

  2001cites this paper
• The murine perilipin gene: the lipid droplet-associated perilipins derive from tissue-specific, mRNA splice variants and define a gene family of ancient origin

  2001cites this paper
• Serial changes in the myocardial beta-adrenergic signalling system in two models of non-insulin dependent diabetes mellitus.

  2001cites this paper
• Structure-function relationships of hormone-sensitive lipase.

  2001cites this paper
• Structure-function relationships of hormone-sensitive lipase.

  2001cites this paper
• Intracellular lipid particles of eukaryotic cells.

  2000cites this paper
• Translocation of Hormone-sensitive Lipase and Perilipin upon Lipolytic Stimulation of Rat Adipocytes*

  2000cites this paper
• Perilipin A Increases Triacylglycerol Storage by Decreasing the Rate of Triacylglycerol Hydrolysis*

  2000cites this paper
• Perilipins, ADRP, and other proteins that associate with intracellular neutral lipid droplets in animal cells.

  1999cites this paper