Acyl Chain Length and Saturation Modulate Interleaflet Coupling in Asymmetric Bilayers: Effects on Dynamics and Structural Order  Salvatore Chiantia,

Slides:



Advertisements
Similar presentations
Probing α-310 Transitions in a Voltage-Sensing S4 Helix
Advertisements

Probing α-310 Transitions in a Voltage-Sensing S4 Helix
Volume 111, Issue 7, Pages (October 2016)
Michiko Tashiro, Hana Inoue, Masato Konishi  Biophysical Journal 
Investigation of Domain Formation in Sphingomyelin/Cholesterol/POPC Mixtures by Fluorescence Resonance Energy Transfer and Monte Carlo Simulations  Monica.
Laurdan Fluorescence Lifetime Discriminates Cholesterol Content from Changes in Fluidity in Living Cell Membranes  Ottavia Golfetto, Elizabeth Hinde,
Madoka Suzuki, Hideaki Fujita, Shin’ichi Ishiwata  Biophysical Journal 
Volume 96, Issue 10, Pages (May 2009)
Volume 84, Issue 4, Pages (April 2003)
Indrajeet Singh, Efrosyni Themistou, Lionel Porcar, Sriram Neelamegham 
Bipedal Locomotion in Crawling Cells
Sphingomyelin/Phosphatidylcholine/Cholesterol Phase Diagram: Boundaries and Composition of Lipid Rafts  Rodrigo F.M. de Almeida, Aleksandre Fedorov, Manuel.
Influence of Chain Length and Unsaturation on Sphingomyelin Bilayers
Volume 113, Issue 12, Pages (December 2017)
Volume 99, Issue 10, Pages (November 2010)
Volume 106, Issue 12, Pages (June 2014)
Membrane Permeability of Hydrocarbon-Cross-Linked Peptides
Volume 112, Issue 7, Pages (April 2017)
Composition Fluctuations in Lipid Bilayers
Hiren Patel, Quang Huynh, Dominik Bärlehner, Heiko Heerklotz 
Juan M. Vanegas, Maria F. Contreras, Roland Faller, Marjorie L. Longo 
Joseph M. Johnson, William J. Betz  Biophysical Journal 
Jason M. Warner, Ben O’Shaughnessy  Biophysical Journal 
Pulsatile Lipid Vesicles under Osmotic Stress
Volume 104, Issue 3, Pages (February 2013)
Modes of Diffusion of Cholera Toxin Bound to GM1 on Live Cell Membrane by Image Mean Square Displacement Analysis  Pierre D.J. Moens, Michelle A. Digman,
Volume 101, Issue 7, Pages (October 2011)
Experimental and Computational Studies Investigating Trehalose Protection of HepG2 Cells from Palmitate-Induced Toxicity  Sukit Leekumjorn, Yifei Wu,
Agata Witkowska, Reinhard Jahn  Biophysical Journal 
Masataka Chiba, Makito Miyazaki, Shin’ichi Ishiwata 
Modulating Vesicle Adhesion by Electric Fields
Xiao-Han Li, Elizabeth Rhoades  Biophysical Journal 
Volume 114, Issue 12, Pages (June 2018)
Fiber-Dependent and -Independent Toxicity of Islet Amyloid Polypeptide
Giant Unilamellar Vesicles Formed by Hybrid Films of Agarose and Lipids Display Altered Mechanical Properties  Rafael B. Lira, Rumiana Dimova, Karin A.
Volume 93, Issue 2, Pages (July 2007)
Yuno Lee, Philip A. Pincus, Changbong Hyeon  Biophysical Journal 
Role of Cholesterol in the Formation and Nature of Lipid Rafts in Planar and Spherical Model Membranes  Jonathan M. Crane, Lukas K. Tamm  Biophysical.
Membrane Elasticity in Giant Vesicles with Fluid Phase Coexistence
Sarah L. Veatch, Sarah L. Keller  Biophysical Journal 
Volume 105, Issue 9, Pages (November 2013)
Volume 110, Issue 3, Pages (February 2016)
Volume 113, Issue 6, Pages (September 2017)
Jonathan P. Litz, Niket Thakkar, Thomas Portet, Sarah L. Keller 
Obstructed Diffusion in Phase-Separated Supported Lipid Bilayers: A Combined Atomic Force Microscopy and Fluorescence Recovery after Photobleaching Approach 
G. Garbès Putzel, Mark J. Uline, Igal Szleifer, M. Schick 
Volume 99, Issue 8, Pages (October 2010)
Volume 95, Issue 9, Pages (November 2008)
Volume 109, Issue 3, Pages (August 2015)
Abhishek Mandal, Patrick C.A. van der Wel  Biophysical Journal 
Lipid Raft Composition Modulates Sphingomyelinase Activity and Ceramide-Induced Membrane Physical Alterations  Liana C. Silva, Anthony H. Futerman, Manuel.
Chang-Chun Lee, Yen Sun, Huey W. Huang  Biophysical Journal 
Saswata Sankar Sarkar, Jayant B. Udgaonkar, Guruswamy Krishnamoorthy 
Cholesterol Modulates the Dimer Interface of the β2-Adrenergic Receptor via Cholesterol Occupancy Sites  Xavier Prasanna, Amitabha Chattopadhyay, Durba.
The Structural Basis of Cholesterol Accessibility in Membranes
Saswata Sankar Sarkar, Jayant B. Udgaonkar, Guruswamy Krishnamoorthy 
Miscibility Critical Pressures in Monolayers of Ternary Lipid Mixtures
Philip J. Robinson, Teresa J.T. Pinheiro  Biophysical Journal 
Volume 98, Issue 1, Pages (January 2010)
Volume 109, Issue 9, Pages (November 2015)
Volume 105, Issue 11, Pages (December 2013)
The Functional Activity of the Human Serotonin 5-HT1A Receptor Is Controlled by Lipid Bilayer Composition  M. Gertrude Gutierrez, Kylee S. Mansfield,
Chze Ling Wee, David Gavaghan, Mark S.P. Sansom  Biophysical Journal 
Phase Equilibria in DOPC/DPPC-d62/Cholesterol Mixtures
Volume 112, Issue 7, Pages (April 2017)
Madoka Suzuki, Hideaki Fujita, Shin’ichi Ishiwata  Biophysical Journal 
Kinetic Folding Mechanism of Erythropoietin
William J. Galush, Jeffrey A. Nye, Jay T. Groves  Biophysical Journal 
Volume 102, Issue 9, Pages (May 2012)
Laurdan Fluorescence Lifetime Discriminates Cholesterol Content from Changes in Fluidity in Living Cell Membranes  Ottavia Golfetto, Elizabeth Hinde,
Presentation transcript:

Acyl Chain Length and Saturation Modulate Interleaflet Coupling in Asymmetric Bilayers: Effects on Dynamics and Structural Order  Salvatore Chiantia, Erwin London  Biophysical Journal  Volume 103, Issue 11, Pages 2311-2319 (December 2012) DOI: 10.1016/j.bpj.2012.10.033 Copyright © 2012 Biophysical Society Terms and Conditions

Figure 1 Interleaflet coupling is influenced by SM acyl chain length. (A) Diffusion coefficient of NBD-DOPE in the inner leaflet as a function of the diffusion coefficient of Atto-SM in the outer leaflet. D values are shown as a percentage of the value measured for the same leaflet localization of the probes in a symmetric DOPC vesicle. Each black circle refers to a single DOPCi/bSMo GUV. Gray triangles refer to single DOPCi/mSMo vesicles. Notice that in all cases shown here, and in later figures, after exchange the outer leaflet contains a mixture of SM and residual inner leaflet PC. The square in position (100, 100) indicates the relative D values measured in a symmetric DOPC GUV. The dash-dotted line has unitary slope (i.e., maximum coupling); the dashed line has null slope (i.e., zero coupling). (B) Box plot of the coupling coefficients measured for each asymmetric GUV in three different types of sample. The median, the lower quartile, and the upper quartile are shown as a line inside, and the lower and upper horizontal boundaries, respectively, of the solid rectangles. The ends of the whiskers represent the 9th and 91st percentiles of the observed data. Two lower outliers for the DOPCi/bSMo sample were removed for presentation. The coupling coefficients are calculated as the slope of the lines connecting each point in A with the (calibration) symmetric GUV in position (100, 100). The coupling coefficients calculated for DOPCi/mSMo vesicles are significantly different from those calculated for the DOPCi/bSMo vesicle, with p < 0.01 measured by Student’s t-test. The numbers of analyzed vesicles were 32 from four independent preparations with bSMo; 24 from three independent preparations with mSMo; 20 from two independent preparations with a 1:1 (mol:mol) mixture of C24:0SM and bSM introduced into the outer leaflet, i.e. (C24:0 + bSM 1:1)o. Independent preparations were produced using different SM/MβCD concentrations, ranging from 42.5 to 57 mM MβCD, in the donor solution. Vesicles with a relative diffusion in the outer leaflet >80% (i.e., 8 μm2/s) were excluded from further analysis and are not represented in the figure. Biophysical Journal 2012 103, 2311-2319DOI: (10.1016/j.bpj.2012.10.033) Copyright © 2012 Biophysical Society Terms and Conditions

Figure 2 Interleaflet coupling is influenced by PC acyl chain saturation. (A) Diffusion coefficient of NBD-DOPE in the inner leaflet as a function of the diffusion coefficient of Atto-SM in the outer leaflet, relative to the value measured with the same configuration of the probes but in a symmetric DOPC or POPC vesicle. Each black circle refers to a single DOPCi/bSMo GUV. Gray triangles refer to POPCi/bSMo vesicles. The point in position (100, 100) indicates the normalized D values measured in a symmetric DOPC or POPC GUV. The dash-dotted line has unitary slope (i.e., maximum expected coupling); the dashed line has null slope (i.e., zero coupling). (B) Box plot of the coupling coefficients measured for each asymmetric GUV with bSM in the outer leaflet and different PCs in the inner leaflet. The coupling coefficients calculated for DOPCi/bSMo vesicles are, as an ensemble, significantly different from those calculated for the POPCi/bSMo vesicle, as measured by Student’s t-test, p < 0.01. The numbers of analyzed vesicles were 32 from four independent preparations with DOPCi; 16 from two independent preparations with POPCi, 22 from two independent preparations with SOPCi; and 22 from two independent preparations with OMPCi. Independent preparations were produced using different MβCD concentrations, ranging from 50 to 57 mM, in the donor solution. Vesicles with a relative diffusion in the outer leaflet >80% were excluded from further analysis and are not represented in the figure. Biophysical Journal 2012 103, 2311-2319DOI: (10.1016/j.bpj.2012.10.033) Copyright © 2012 Biophysical Society Terms and Conditions

Figure 3 Probing the state of the inner leaflet using the fluorescence lifetime of NBD-DPPE in asymmetric GUVs and SUVs. (A) Fast fluorescence lifetime component of NBD-DPPE (τ1) in symmetric and asymmetric GUVs as calculated from FLIM. At least six images from two independent preparations were analyzed for each lipid composition (see Fig. S3). τ1 measured for the inner leaflet of symmetric DOPC GUVs is significantly different from τ1 measured for both leaflets of asymmetric DOPCi/bSMo GUVs (t-test p < 0.05). τ1 measured for the inner leaflet of symmetric POPC GUVs is significantly different from τ1 measured for both leaflets of asymmetric POPCi/bSMo GUVs (t-test p < 0.01). The asterisk indicates data sets statistically not distinguishable by t-test. Error bars represent standard deviations. (B) Short fluorescence lifetime component of NBD-DPPE τ1 in asymmetric SUVs of varying composition. Lines are reference values for symmetric DOPC inner leaflet (dash-dotted line), symmetric POPC inner leaflet (dashed line), symmetric DOPC/bSM 1:4 inner leaflet (dash-dot-dotted line) and symmetric POPC/bSM 1:4 inner leaflet (short-dash-long-dashed line). All the reported values were obtained from four measurements in two independent preparations. Error bars represent the standard deviations. Biophysical Journal 2012 103, 2311-2319DOI: (10.1016/j.bpj.2012.10.033) Copyright © 2012 Biophysical Society Terms and Conditions

Figure 4 Interleaflet coupling is not detected by either TMA-DPH anisotropy or NR12S emission wavelength. (A) Maximum emission wavelength of NR12S in the inner leaflet of asymmetric SUVs with varying composition. Lines are reference values for symmetric DOPC inner leaflet (dash-dotted line), and symmetric DOPC/bSM 1:4 inner leaflet (dash-dot-dotted line). Reference values for symmetric POPC and symmetric POPC/bSM 1:4 are within ±1 nm of those indicated for symmetric DOPC and symmetric DOPC/bSM 1:4, respectively. All the reported values were obtained from four measurements in two independent preparations, and the error bars represent standard deviations. (B) TMA-DPH anisotropy measured either in the outer (gray) or inner leaflet (black) of asymmetric SUVs with varying composition. Lines are reference values for symmetric DOPC (dash-dotted line) and symmetric DOPC/bSM 1:4 (dash-dot-dotted line). Reference values for symmetric POPC and symmetric POPC/bSM 1:4 are within ±0.01 of those indicated for symmetric DOPC and symmetric DOPC/bSM 1:4, respectively. All the reported values were obtained from four measurements in two independent preparations, and the error bars represent standard deviations. Biophysical Journal 2012 103, 2311-2319DOI: (10.1016/j.bpj.2012.10.033) Copyright © 2012 Biophysical Society Terms and Conditions

Figure 5 A model for how acyl chain composition influences interleaflet coupling. Schematic models of asymmetric bilayers with inner leaflets composed of DOPC (blue) (A and B) or a partially saturated PC, i.e., POPC, SOPC, or OMPC (red) (C and D). The outer leaflet is composed in all cases of SM (mSM (B) and bSM (A, C, and D)). The green rectangles indicate regions near the bilayer midplane, where acyl chains from an opposing leaflet might interact. Red lines represent the long interdigitating acyl chains of mSM in B, and the saturated PC acyl chains in C and D. Biophysical Journal 2012 103, 2311-2319DOI: (10.1016/j.bpj.2012.10.033) Copyright © 2012 Biophysical Society Terms and Conditions