title: Activation of the lipid droplet controls the rate of lipolysis of triglycerides in the insect fat body.
abstract: The hydrolysis of triglyceride (TG) stored in the lipid droplets of the insect fat body is under hormonal regulation by the adipokinetic hormone (AKH), which triggers a rapid activation cAMP-dependent kinase cascade (protein kinase A (PKA)). The role of phosphorylation on two components of the lipolytic process, the TG-lipase and the lipid droplet, was investigated in fat body adipocytes. The activity of purified TG-lipase determined using in vivo TG-radiolabeled lipid droplets was unaffected by the phosphorylation of the lipase. However, the activity of purified lipase was 2.4-fold higher against lipid droplets isolated from hormone-stimulated fat bodies than against lipid droplets isolated from unstimulated tissue. In vivo stimulation of lipolysis promotes a rapid phosphorylation of a lipid droplet protein with an apparent mass of 42-44 kDa. This protein was identified as "Lipid Storage Droplet Protein 1" (Lsdp1). In vivo phosphorylation of this protein reached a peak approximately 10 min after the injection of AKH. Supporting a role of Lsdp1 in lipolysis, maximum TG-lipase activity was also observed with lipid droplets isolated 10 min after hormonal stimulation. The activation of lipolysis was reconstituted in vitro using purified insect PKA and TG-lipase and lipid droplets. In vitro phosphorylation of lipid droplets catalyzed by PKA enhanced the phosphorylation of Lsdp1 and the lipolytic rate of the lipase, demonstrating a prominent role PKA and protein phosphorylation on the activation of the lipid droplets. AKH-induced changes in the properties of the substrate do not promote a tight association of the lipase with the lipid droplets. It is concluded that the lipolysis in fat body adipocytes is controlled by the activation of the lipid droplet. This activation is achieved by PKA-mediated phosphorylation of the lipid droplet. Lsdp1 is the main target of PKA, suggesting that this protein is a major player in the activation of lipolysis in insects.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, 74078, USA.
authors.forename: Rajesh T
authors.initials: RT
authors.lastname: Patel
authors.forename: Jose L
authors.initials: JL
authors.lastname: Soulages
authors.forename: Balaji
authors.initials: B
authors.lastname: Hariharasundaram
authors.forename: Estela L
authors.initials: EL
authors.lastname: Arrese
mesh_terms: D017667:Adipocytes / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins; D002384:Catalysis; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007301:Insect Hormones / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry / Q000502:physiology; D007313:Insecta; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D008066:Lipolysis; D018402:Manduca; D013058:Mass Spectrometry; D009842:Oligopeptides / Q000737:chemistry; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000737:chemistry* / Q000502:physiology*; D010766:Phosphorylation; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D014280:Triglycerides / Q000737:chemistry*
chemical_list: D002352:Carrier Proteins; D007301:Insect Hormones; D019476:Insect Proteins; D008055:Lipids; D009842:Oligopeptides; D000070778:Perilipin-1; D010750:Phosphoproteins; D014280:Triglycerides; C044833:adipokinetic hormone; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D011761:Pyrrolidonecarboxylic Acid