title: Membrane attachment and structure models of lipid storage droplet protein 1.
abstract: Neutral lipid triglycerides, a main reserve for fat and energy, are stored in organelles called lipid droplets. The storage and release of triglycerides are actively regulated by several proteins specific to the droplet surface, one of which in insects is PLIN1. PLIN1 plays a key role in the activation of triglyceride hydrolysis upon phosphorylation. However, the structure of PLIN1 and its relation to functions remain elusive due to its insolubility and crystallization difficulty. Here we report the first solid-state NMR study on the Drosophila melanogaster PLIN1 in combination with molecular dynamics simulation to show the structural basis for its lipid droplet attachment. NMR spin diffusion experiments were consistent with the predicted membrane attachment motif of PLIN1. The data indicated that PLIN1 has close contact with the terminal methyl groups of the phospholipid acyl chains. Structure models for the membrane attachment motif were generated based on hydrophobicity analysis and NMR membrane insertion depth information. Simulated NMR spectra from a trans-model agreed with experimental spectra. In this model, lipids from the bottom leaflet were very close to the surface in the region enclosed by membrane attachment motif. This may imply that in real lipid droplet, triglyceride molecules might be brought close to the surface by the same mechanism, ready to leave the droplet in the event of lipolysis. Juxtaposition of triglyceride lipase structure to the trans-model suggested a possible interaction of a conserved segment with the lipase by electrostatic interactions, opening the lipase lid to expose the catalytic center.
authors: Department of Physics, 230 L Henry Bellmon Research Center, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Penghui
authors: P
authors: Lin
authors: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Xiao
authors: X
authors: Chen
authors: Department of Physics, 230 L Henry Bellmon Research Center, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Hem
authors: H
authors: Moktan
authors: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Estela L
authors: EL
authors: Arrese
authors: Department of Physics, 230 L Henry Bellmon Research Center, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Lian
authors: L
authors: Duan
authors: Department of Physics, 230 L Henry Bellmon Research Center, Oklahoma State University, Stillwater, OK 74078, USA; State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan Center for Magnetic Resonance, Key Laboratory of Magnetic Resonance in Biological Systems, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071, China.
authors: Liying
authors: L
authors: Wang
authors: Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
authors: Jose L
authors: JL
authors: Soulages
authors: Department of Physics, 230 L Henry Bellmon Research Center, Oklahoma State University, Stillwater, OK 74078, USA. Electronic address: donghua@okstate.edu.
authors: Donghua H
authors: DH
authors: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008563:Membrane Lipids / Q000378:metabolism*; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013917:Thrombin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D008074:Lipoproteins; D008563:Membrane Lipids; D000070778:Perilipin-1; D010750:Phosphoproteins; D013917:Thrombin
keywords: 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol; CP; DARR; DMPG; Lipid storage droplet protein; Lsd1; MAS; MD; MD simulation; Magic-angle spinning; PAT family; Proton spin diffusion; Solid-state NMR; Triglyceride lipolysis; cross polarization; dipolar-assisted rotational resonance; lipid storage droplet protein 1; magic-angle spinning; molecular dynamics; perilipin, ADRP, TIP 47 family