Phosphatidylinositol-4,5-bisphosphate ionization and domain formation in the presence of lipids with hydrogen bond donor capabilities

Graber, Zachary T.; Jiang, Zhiping; Gericke, Arne; Kooijman, Edgar E.

doi:10.1016/j.chemphyslip.2012.07.003

Cited by 43 publications

(56 citation statements)

References 21 publications

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“…However, assuming an effective charge between Ϫ3e and Ϫ4e for PI(4,5)P 2 at physiological pH (66), the charge density of a membrane surface containing 5 mol% PI(4,5)P 2 is lower than that brought about by 30 mol% DOPS. The experimental result then shows that PI(4,5)P 2 binding to MA involves more than simple electrostatic interaction.…”

Section: Discussionmentioning

confidence: 99%

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: 99%

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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“…The binding of PLCβ1 and tubulation (22 ± 4%; t test compared between with EDTA and without CaCl 2 , P > 0.05) were recovered by the addition of Ca 2+ chelating reagents. The surface charge of lipid head groups was affected by the basic pH (21,22). PIP 2 ionization and domain formation were reported in the presence of lipids with hydrogen bond donor capabilities (21,23).…”

Section: +mentioning

confidence: 99%

“…The surface charge of lipid head groups was affected by the basic pH (21,22). PIP 2 ionization and domain formation were reported in the presence of lipids with hydrogen bond donor capabilities (21,23). We also analyzed the effect of neutral pH on PLCβ1b-induced tubulation.…”

Section: +mentioning

confidence: 99%

Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation

Inaba

Kishimoto

Murate

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Lipid membrane curvature plays important roles in various physiological phenomena. Curvature-regulated dynamic membrane remodeling is achieved by the interaction between lipids and proteins. So far, several membrane sensing/sculpting proteins, such as Bin/ amphiphysin/Rvs (BAR) proteins, are reported, but there remains the possibility of the existence of unidentified membrane-deforming proteins that have not been uncovered by sequence homology. To identify new lipid membrane deformation proteins, we applied liposome-based microscopic screening, using unbiased-darkfield microscopy. Using this method, we identified phospholipase Cβ1 (PLCβ1) as a new candidate. PLCβ1 is well characterized as an enzyme catalyzing the hydrolysis of phosphatidylinositol-4,5-bisphosphate (PIP 2 ). In addition to lipase activity, our results indicate that PLCβ1 possessed the ability of membrane tubulation. Lipase domains and inositol phospholipids binding the pleckstrin homology (PH) domain of PLCβ1 were not involved, but the C-terminal sequence was responsible for this tubulation activity. Computational modeling revealed that the C terminus displays the structural homology to the BAR domains, which is well known as a membrane sensing/sculpting domain. Overexpression of PLCβ1 caused plasma membrane tubulation, whereas knockdown of the protein reduced the number of caveolae and induced the evagination of caveolin-rich membrane domains. Taken together, our results suggest a new function of PLCβ1: plasma membrane remodeling, and in particular, caveolae formation.phospholipase Cβ1 | membrane tubulation | microscopy screening | caveolae | BAR-like domain

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“…Here we used a system containing 20% PS and 3% di-oleoyl-phosphatidylinositol-(4,5)-bisphosphate. The latter lipid contains a negative charge of 4 at physiological pH of 7.4 [9,38,39]. Thus, the overall surface charge of SUVs composed of POPC/DOPC/Chol/POPS/PI(4,5)P 2 (37:20: 20:20:3) is very high, which may hinder vesicle adsorption or rupture on a support.…”

Section: Discussionmentioning

confidence: 98%

Importance of phospholipid bilayer integrity in the analysis of protein–lipid interactions

Drücker

Gerke

Galla

2014

Biochemical and Biophysical Research Communications

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Phosphatidylinositol-4,5-bisphosphate ionization and domain formation in the presence of lipids with hydrogen bond donor capabilities

Cited by 43 publications

References 21 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

Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation

Importance of phospholipid bilayer integrity in the analysis of protein–lipid interactions

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