Influence of Annexin V on the Structure and Dynamics of Phosphatidylcholine/Phosphatidylserine Bilayers:  A Fluorescence and NMR Study

Saurel, Olivier; Cézanne, Laurence; Milon, Alain; Tocanne, Jean‐François; Demange, Pascal

doi:10.1021/bi971484n

Cited by 53 publications

(69 citation statements)

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“…A slight ordering in the DMPA component above its phase transition, in the presence of protein, is suggested from the reduction in frequency at temperatures above the completion of the melting process. This minor change is consistent with peripheral protein binding and with a recent study of Saurel et al (12), who examined the consequences of AxV binding on the structure and dynamics of PC/PS bilayers using a variety of fluorescence and NMR-based experiments. They observed the appearance of slow motions (on the NMR time scale) in the lipid spectra, indicative of a decrease in lipid lateral diffusion, but observed no indication of changes in the lipid molecular packing and acyl chain flexibility in either of the lipid components.…”

Section: Discussionsupporting

confidence: 88%

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Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

Flach

Seaton

et al. 1998

Biochemistry

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Annexin V (AxV) is a member of a family of proteins that exhibit functionally relevant Ca2+-dependent binding to anionic phospholipid membranes. Protein structure and stability as a function of Ca2+ and phospholipids was studied by bulk phase infrared (IR) spectroscopy and by IR reflection-absorption spectroscopy (IRRAS) of monolayers in situ at the air/water (A/W) interface. Bulk phase experiments revealed that AxV undergoes an irreversible thermal denaturation at approximately 45-50 degreesC, as shown by the appearance of amide I bands at 1617 and 1682 cm-1. However, some native secondary structure is retained, even at 60 degreesC, consistent with a partially unfolded "molten globule" state. Formation of the Ca2+/phospholipid/protein ternary complex significantly protects the protein from thermal denaturation as compared to AxV alone, Ca2+/AxV, or lipid/AxV mixtures. Stabilization of AxV secondary structure by a DMPA monolayer in the presence of Ca2+ was also observed by IRRAS. Spectra of an adsorbed AxV film in the presence or absence of Ca2+ showed a 10 cm-1 shift in the amide I mode, corresponding to loss of ordered structure at the A/W interface. In both the bulk phase and IRRAS experiments, protection against H-->D exchange in AxV was enhanced only in the ternary complex. The combined data suggest that the secondary structure of AxV is strongly affected by the Ca2+/membrane component of the ternary complex whereas lipid conformational order is unchanged by protein.

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Section: Discussionsupporting

confidence: 88%

“…The effect of protein binding on the structure of mixed PC/PS vesicles and oriented bilayers was studied by fluorescence and NMR techniques (12). The presence of AxV led to the appearance of slow motions on the NMR time scale, indicating a reduction in the lipid diffusion rate.…”

mentioning

confidence: 99%

Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

Flach

Seaton

et al. 1998

Biochemistry

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“…This also shows that the solution in the thin gap (∼ 8 nm for the 24-mer hybrid and ∼ 16 nm for the 48-mer hybrid, see below) between the bilayer patch and substrate surface is readily water-permeable so that salt ions are washed out immediately. Water accessibility was also checked by a cobalt quenching experiment (Saurel, Cezanne et al 1998). Co 2+ completely quenches fluorescence from Texas red labeled lipid, and in the case of solid supported lipid membranes, 54% of the fluorescence intensity is immediately quenched because Co 2+ in bulk solution cannot access the fluorescence dye in the bottom leaflet close to the substrate (Ajo-Franklin, Yoshina-Ishii et al 2005).…”

Section: Resultsmentioning

confidence: 99%

DNA-tethered membranes formed by giant vesicle rupture

Chung

Lowe

Chan

et al. 2009

Journal of Structural Biology

105

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We have developed a strategy for preparing tethered lipid bilayer membrane patches on solid surfaces by DNA hybridization. In this way, the tethered membrane patch is held at a controllable distance from the surface by varying the length of the DNA used. Two basic strategies are described. In the first, single-stranded DNA strands are immobilized by click chemistry to a silica surface, whose remaining surface is passivated to prevent direct assembly of a solid supported bilayer. Then giant unilamellar vesicles (GUVs) displaying the antisense strand, using a DNA-lipid conjugate developed in earlier work (Chan, Lengerich et al. 2008), are allowed to tether, spread and rupture to form tethered bilayer patches. In the second, a supported lipid bilayer displaying DNA using the DNA-lipid conjugate is first assembled on the surface. Then GUVs displaying the antisense strand are allowed to tether, spread and rupture to form tethered bilayer patches. The essential difference between these methods is that the tethering hybrid DNA is immobile in the first, while it is mobile in the second. Both strategies are successful; however, with mobile DNA hybrids as tethers, the patches are unstable, while in the first strategy stable patches can be formed. In the case of mobile tethers, if different length DNA hybrids are present, lateral segregation by length occurs and can be visualized by fluorescence interference contrast microscopy making this an interesting model for interactions that occur in cell junctions. In both cases, lipid mobility is high and there is a negligible immobile fraction. Thus, these architectures offer a flexible platform for the assembly of lipid bilayers at a well-defined distance from a solid support. KeywordsTethered lipid membrane; surface-immobilized DNA; Giant unilamellar vesicles Supported lipid bilayers (SLBs) have been widely used as a model for cell membranes (Sackmann 1996;Chan and Boxer 2007) and to investigate membrane components including proteins in a simpler context apart from the complex cellular environment. SLBs are assembled by Langmuir-Blodgett techniques or spontaneous fusion of unilamellar vesicles on carefully prepared surfaces, usually hydrophilic solid supports, such as glass (Seu, Pandey et al. 2007), silica, mica (Richter, Berat et al. 2006), or TiO 2 (Rossetti, Bally et al. 2005). Although SLBs have the advantages of simple formation, easy handling and are well-suited for investigation by a suite of surface sensitive methods due to their planar geometry, the close proximity of the lower leaflet to the solid support often leads to unfavorable interactions with integral membrane proteins. Recognizing this limitation, many groups have described methods to separate the © 2009 Elsevier Inc. All rights reserved. *Corresponding Author: Tel.: 1-650-723-4482; Fax: 1-650-723-4817; E-mail: sboxer@stanford.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early versi...

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“…Several annexin gene products have been shown to cause pronounced effects on lipid dynamics after Ca 2ϩ -dependent membrane binding. FRAP experiments have shown that the lateral diffusion rates of phospholipids are reduced by several orders of magnitude by annexin A4 or A5 binding (29,30). Furthermore, studies of spin-labeled lipids show that annexin A5 has found profound effects on the lipid fluidity gradient (31,32), which is naturally present in phospholipid bilayers (25).…”

Section: Discussionmentioning

confidence: 99%

Structure and Dynamics of a Helical Hairpin that Mediates Calcium-dependent Membrane Binding of Annexin B12

Isas

Langen

Hubbell

et al. 2004

Journal of Biological Chemistry

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A wealth of high-resolution structural data has accumulated for soluble annexins, but only limited information is available for the biologically important membrane-bound proteins. To investigate the structural and dynamic changes that occur upon membrane binding, we analyzed the electron paramagnetic resonance (EPR) mobility and accessibility parameters of a contin- showed that the overall backbone fold for the scanned region did not change upon membrane binding. However, side-chains in the loop between the D and E helices were highly dynamic in solution but became essentially frozen in the EPR time scale upon binding to membranes. Accessibility measurements clearly established that side-chains in this loop were exposed to the hydrophobic core of the bilayer and provide the first evidence that a D-E loop directly participates in the Ca 2؉ -dependent binding of annexins to membranes. Other localized changes showed that the D-helix became much less dynamic after membrane binding and identified quaternary contact sites in the membrane-bound homo-trimer. Finally, immobilization of the D-E loop upon contact with phospholipid suggests that the bilayer, which is normally very mobile on the EPR time scale, is immobilized in the head-group region by the annexin B12. This suggests that annexin B12 alters membrane structure in a manner that may be biologically significant.

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Influence of Annexin V on the Structure and Dynamics of Phosphatidylcholine/Phosphatidylserine Bilayers: A Fluorescence and NMR Study

Cited by 53 publications

References 50 publications

Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

DNA-tethered membranes formed by giant vesicle rupture

Structure and Dynamics of a Helical Hairpin that Mediates Calcium-dependent Membrane Binding of Annexin B12

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Influence of Annexin V on the Structure and Dynamics of Phosphatidylcholine/Phosphatidylserine Bilayers: A Fluorescence and NMR Study

Cited by 53 publications

References 50 publications

Stability of Annexin V in Ternary Complexes with Ca2+ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

Stability of Annexin V in Ternary Complexes with Ca2+ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

DNA-tethered membranes formed by giant vesicle rupture

Structure and Dynamics of a Helical Hairpin that Mediates Calcium-dependent Membrane Binding of Annexin B12

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Product

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Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases

Stability of Annexin V in Ternary Complexes with Ca²⁺ and Anionic Phospholipids: IR Studies of Monolayer and Bulk Phases