Myristoylation Restricts Orientation of the GRASP Domain on Membranes and Promotes Membrane Tethering

Heinrich, Frank; Nanda, Hirsh; Goh, Haw Zan; Bachert, Collin; Lösche, Mathias; Linstedt, Adam D.

doi:10.1074/jbc.m113.543561

Cited by 36 publications

(58 citation statements)

References 41 publications

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“…(ii) The myristoyl group may only partially insert, as has been previously proposed in a different molecular context (60). (iii) A myristoyl membrane anchor further reduces the bound protein's entropy compared to the nonlipidated species, as recently shown for the membrane-bound, myristoylated GRASP domain (62).…”

Section: Discussionmentioning

confidence: 75%

Membrane Binding of HIV-1 Matrix Protein: Dependence on Bilayer Composition and Protein Lipidation

Barros

Heinrich

Datta

et al. 2016

J Virol

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By assembling in a protein lattice on the host's plasma membrane, the retroviral Gag polyprotein triggers formation of the viral protein/membrane shell. The MA domain of Gag employs multiple signals-electrostatic, hydrophobic, and lipid-specific-to bring the protein to the plasma membrane, thereby complementing protein-protein interactions, located in full-length Gag, in lattice formation. We report the interaction of myristoylated and unmyristoylated HIV-1 Gag MA domains with bilayers composed of purified lipid components to dissect these complex membrane signals and quantify their contributions to the overall interaction. Surface plasmon resonance on well-defined planar membrane models is used to quantify binding affinities and amounts of protein and yields free binding energy contributions, ⌬G, of the various signals. Charge-charge interactions in the absence of the phosphatidylinositide PI(4,5)P 2 attract the protein to acidic membrane surfaces, and myristoylation increases the affinity by a factor of 10; thus, our data do not provide evidence for a PI(4,5)P 2 trigger of myristate exposure. Lipid-specific interactions with PI(4,5)P 2 , the major signal lipid in the inner plasma membrane, increase membrane attraction at a level similar to that of protein lipidation. While cholesterol does not directly engage in interactions, it augments protein affinity strongly by facilitating efficient myristate insertion and PI(4,5)P 2 binding. We thus observe that the isolated MA protein, in the absence of protein-protein interaction conferred by the full-length Gag, binds the membrane with submicromolar affinities. IMPORTANCELike other retroviral species, the Gag polyprotein of HIV-1 contains three major domains: the N-terminal, myristoylated MA domain that targets the protein to the plasma membrane of the host; a central capsid-forming domain; and the C-terminal, genome-binding nucleocapsid domain. These domains act in concert to condense Gag into a membrane-bounded protein lattice that recruits genomic RNA into the virus and forms the shell of a budding immature viral capsid. In binding studies of HIV-1 Gag MA to model membranes with well-controlled lipid composition, we dissect the multiple interactions of the MA domain with its target membrane. This results in a detailed understanding of the thermodynamic aspects that determine membrane association, preferential lipid recruitment to the viral shell, and those aspects of Gag assembly into the membrane-bound protein lattice that are determined by MA. T he polyprotein Gag is an essential component for the reproduction of retroviruses, such as HIV-1, in infected cells. In the production of new viruses, many copies of Gag assemble laterally, either in the cytoplasm or on the plasma membrane (PM) of the host, thus acquiring their lipid envelope as they bud from the cell. Distinct retroviral genera encode Gag proteins which all show a similar domain architecture, with an N-terminal matrix (MA) domain that targets the PM, followed by the capsid (CA) and nucleocapsid ...

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Section: Discussionmentioning

confidence: 75%

Membrane Binding of HIV-1 Matrix Protein: Dependence on Bilayer Composition and Protein Lipidation

Barros

Heinrich

Datta

et al. 2016

J Virol

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“…3A (Table 1). All measurements were evaluated using a global fit to obtain a structural profile of the stBLM and of the enzyme at the membrane interface (28,31). The resulting bilayer structure and median protein envelope yield an excellent fit to the measured reflectivity (Fig.…”

Section: Effect Of ␣-Synuclein On Gcase Membrane Binding Probed Bymentioning

confidence: 99%

“…1A), a portion of the enzyme interacts within the phospholipid headgroups and penetrates into the acyl chain region of the outer leaflet. Using previously established rigid body modeling methods (28,31) and a Monte Carlo Markov chain data refinement algorithm (42), the spatial association and orientation of glycosylated GCase on the stBLM is obtained based on the GCase x-ray crystal structure (Protein Data Bank code 1OGS) (46). Within the 68% probability bounds (Fig.…”

Section: Effect Of ␣-Synuclein On Gcase Membrane Binding Probed Bymentioning

confidence: 99%

“…Data modeling that determines the orientation of a protein at the lipid bilayer requires the use of a structural model that is assumed to be invariant (rigid body modeling) (31,33). For GCase, the Protein Data Bank structure with the identifier 1OGS (46) was augmented by adding hydrogen atoms using the MolProbity Web service (47).…”

Section: ϫ2mentioning

confidence: 99%

“…Although it is of lower resolution than some structural methods, such as x-ray crystallography and solidstate nuclear magnetic resonance spectroscopy, NR has been successfully used to characterize a number of peripheral (e.g. ␣-syn (34), HIV-1 matrix (33), colicin N (35), PTEN tumor suppressor (30), and the GRASP Golgi stacking protein (31)) and integral membrane proteins (e.g. hemolysin (32) and outer membrane protein F (35)) on physiologically relevant, fluid phospholipid bilayers.…”

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Structural Features of Membrane-bound Glucocerebrosidase and α-Synuclein Probed by Neutron Reflectometry and Fluorescence Spectroscopy

Yap¹,

Jiang²,

Heinrich

et al. 2015

Journal of Biological Chemistry

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Background: A specific interaction exists between ␣-synuclein and glucocerebrosidase on the lipid membrane, resulting in enzyme inhibition. Results: Binding glucocerebrosidase has a profound effect on ␣-synuclein, moving roughly half of its embedded helical region above the membrane plane. Conclusion: A model is proposed with structural insights into glucocerebrosidase inhibition by ␣-synuclein. Significance: ␣-Synuclein-glucocerebrosidase interaction provides a molecular connection between Parkinson and Gaucher diseases.

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Golgi Structure and Function in Health, Stress, and Diseases

Ahat

Wang

2019

Results and Problems in Cell Differentiation

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The Golgi apparatus is a central intracellular membrane-bound organelle with key functions in trafficking, processing, and sorting of newly synthesized membrane and secretory proteins and lipids. To best perform these functions, Golgi membranes form a unique stacked structure. The Golgi structure is dynamic but tightly regulated; it undergoes rapid disassembly and reassembly during the cell cycle of mammalian cells and is disrupted under certain stress and pathological conditions. In the past decade, significant amount of effort has been made to reveal the molecular mechanisms that regulate the Golgi membrane architecture and function. Here we review the major discoveries in the mechanisms of Golgi structure formation, regulation, and alteration in relation to its functions in physiological and pathological conditions to further our understanding of Golgi structure and function in health and diseases.

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Myristoylation Restricts Orientation of the GRASP Domain on Membranes and Promotes Membrane Tethering

Cited by 36 publications

References 41 publications

Membrane Binding of HIV-1 Matrix Protein: Dependence on Bilayer Composition and Protein Lipidation

Membrane Binding of HIV-1 Matrix Protein: Dependence on Bilayer Composition and Protein Lipidation

Structural Features of Membrane-bound Glucocerebrosidase and α-Synuclein Probed by Neutron Reflectometry and Fluorescence Spectroscopy

Golgi Structure and Function in Health, Stress, and Diseases

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