Nathaly Marn Medina 03/08/13 Biophysics Seminar

Lipid bilayers
Membrane bilayer
5 - 8 nm thick

Erythrocyte Principles of Biochemistry - Lehninger

Modified from: bioquimicafosfo.blogspot.com

Lipid

Lipids
LIPIDS IN BIOMEMBRANE Semi-permeable barrier Fission/Fusion Tuning proteins function Gating of channels Chemical environment Adjusting membrane curvature

Studying mechanical properties of lipid bilayers

Vesicles under stress Micropipette aspiration Quantitative study of elastic moduli

Mesoscopic outlook
Rawicz et al. Effect of Chain Length and Unsaturation of Elasticity of Lipid Bilayers. 2000 - Biophysical Journal

Atomic Force Microscope (AFM) Topology of lipid bilayers Force spectroscopy mode

Nanometer scale

Atomic Force Microscope

Mirror Laser

Quad PD

Cantilever

Atomic Force Microscope

Mirror Laser

Quad PD

Cantilever

Atomic Force Microscope

PhotoDetector Light source

Focusing optics

Piezo Cantilever Sample

Force spectroscopy with AFM

Indentation on a lipid bilayer

Interaction forces tip-phospholipids

Effect of ionic strength

DMPC bilayer (a) Without NaCl + MgCL2 (b) With NaCl + MgCL2

Effect of temperature
19C 27.2C 30.3C 31.3C 37.5C Gel-fluid phase transition in a DMPC bilayer Gel phase solid ordered phase Melting temperature: ~24C Fluid phase Liquid disordered phase

Effect of length of tails

The longer the apolar chain, the higher the force required to indent the membrane

Conclusions
AFM force spectroscopy New tool to explore the mechanical properties of lipid bilayers Breakthrough force molecular fingerprint Nanometer and piconewton resolution Bridge the gap with MD simulations Chemistry of the phospholipid headgroups Effect on the mechanical stability of the membrane? Fingerprint the mechanical stability of a full cellular

membrane (very ambitious)

Membrane proteins

Complex mixture of phospholipids

Role of each individual phospolipid and protein on the mechanical properties of the membrane

LA COPA DE VINO NOS ESPERA

Effect of variety of phospholipids in the bilayer

Chianta et al. (2006) Phase separation in a raft-exhibiting DOPC/SM/Chol mixture Force required to indent the bilayer

Liquid ordered phase 10.2 nN Liquid disordered phase 6.5 nN

http://www.lanl.gov/science/1663/august2011/story3full.shtml

Sullan et al. (2009) DOPC/SM/Chol + ceramide increases its mechanical stability in both phases Picas et al. (2009) POPE/POPG (3:1) two different calcium-induced domains Higher domains higher mechanical stability (0.92 nN) gel phase Lower domains lower mechanical stability (0.24 nN) fluid phase

Breakthrough force and friction interrelation

Grant and Tiberg (2002) Friction properties of DOPC Resistance to normal loads Efficient role as a lubricant Benz et al. (2004) Friction properties of DPPE/DLPE Single defects in lipid bilayers (AFM) Stability of the bilayer (SFA) Trunfio-Sfarghiu et al. (2008) Bilayers exhibiting a stronger mechanical resistance to indentation lower and more stable friction coefficients Oncins et al. (2005) DMPC bilayer in NaCl buffer solution The presence of Na+ cations induced structural changes in the bilayer Three different friction regimes as the vertical force increased.

Models of film rupture in lipid bilayers

Formation of a hole under the tip Continuum nucleation model Distribution of forces to create a hole is connected to line tension Free energy associated with the unsaturated bonds Molecular model Each molecule has certain energetically favorable binding sites Film pressed by an AFM tip forming a hole is energetically ok These theories represent well the experimental data