Systems

Study Collective Dynamics of Using to Biological and Model Membrane

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 1

June 25-29, 2007 NIST Gaithersburg,

NCNR Summer School, Maikel C. Rheinstädter University of Missouri –University Columbia Department of Physics Astronomy and Organism organization will be considered, ) Organ al methods and concepts … primary goal … the

or purpose differentiates biology from other biology from other differentiates or purpose Hartwell et al. structure and function by application of the and structure :

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“Although living systems obey the laws of physics and chemistry, the the laws of physics and chemistry, living systems obey “Although notion of natural sciences” ( c

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l

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i

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Biological Physics is… f Functional Module

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h c

Biological Macromolecule ar Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 2

as will both experimental and theoretical studies” “the study of biological phenomena using physic of biological is to advance the understanding principles of physical science” “… biophysical approa distinctively a

er

i H Molecule Atom

Aims and Scope, European Biophysical Journal: : D. Neumann, Saarbrücken, Germany Saarbrücken, Neumann, D. Applications Drug transport of physiological Bioengineering: Biosensors using solid supported membranes and biological functionalities: Understanding The Cell Membrane (inter)action

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 3 Membrane is the primary site of on a molecular scale? How can we study structure and dynamics H.Heller, München, Germany Membrane Dynamics

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 4 -> Why can’t we see atoms or molecules? Optical Microscope Optical Techniques

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 5 Magnifying Glass 2 1 v − 2 2 v () m 2 1

‘reciprocal space’ =

ω h energy

π d 2 =

θ

λ sin move.”

π 2 how they r Scattering laws v 4 “…where the − atoms are and = Scattering vector 1 r v q () Fourier Transformation h m = r q Scattering-Reciprocal Space

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 6 momentum

‘real space’ diffraction 2 1.8 1.6 f 1.4 1.2 1 ] g 0.8 -1 [A || q 0.6 Reciprocal Space of a Membrane Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 7 0.4 0.2 0 -0.2 -0.4

1

1.8 1.6 1.4 1.2 ⊥ 0.8 0.6 0.4 0.2

] [A q -1 ] -1 Complex Membrane [Å y q

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 8 -0,2 -0,1 0,0 0,1 0,2 DMPC / Alamethicin = 1:25 T = 22,5°C

0,8 0,7 0,6 0,5 0,4 0,3 0,2 0,1

z

] [Å q -1 Terry P. Orlando move.” how they “…where the atoms are and (k) ω Collective Dynamics - Phonons

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 9 Terry P. Orlando, MIT Dispersion Relation

t (ps) (ps) t t -1 0 1 2 3 4 5 6 10 10 10 10 10 10 10 10 1 10 0 10 0 10 1 10 ) -1 Time-of-Flight (IN5) Time-of-Flight dynamics Q (Å Q R (Å) -1 10 2 Triple-Axis Triple-Axis (IN12/IN8) 10 Backscattering Backscattering IN10/IN16 wave vector resolved access to -Echo (IN11/IN15) Inelastic neutron scattering gives neutron scattering Inelastic -2 10 2 1 0 -1 -2 -3 -4 -5

10 10 10

10 10 10 10 10 E (meV) (meV) E E specific motions

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 10 “Broadband” excitations relaxations 60 Å fluctuations 100 Å Thermal membrane membrane Thermal Mode ++

s e ci n Propagating Oscillating Relaxating e u q re f on ti a s it te xc ra e on of ti a ce x n a e l ‘Phonons in d re membranes’ n e d p n e a -d q

excitations Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 11 Mesoscopic Membrane Fluctuations ) Elementary -1

q (Å

) (ns

τ -1 -1 Dispersion relation Contains ‘dynamic’ information Triple-Axis- Backscattering-Spectrometer || q s μ <1 τ Relaxation Propagation <5000Å & 1ps< ║ DMPC –d54 /q π 2Å<2 Oscillation& Relaxation

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 12 Collective Excitations in model membranes Spectrometer Neutron-Spin-Echo- The ‘Neutron Window’ Phases dynamics and Handbook of Biological Physics Biological Handbook of Lamellar structure Functionality determined by Lipid/Water Lipid/Water Parameters: temperature + hydration fluid =21.5 °C c T DMPC –d54 bilayers gel Spontaneous aggregation to aggregation Spontaneous

(Phospho-)Lipids and Membranes Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 13 tail head Molekulardynamik Heller et al., J-Phys.Chem 93 „Main” transition Hydrophilic Hydrophobic ’ β α P L Simple model system Towards biological membranes

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 14 excitations scattering Collective • in- Coherent quasielastic and neutron • Inelastic X-ray scattering responsible for ‘functionalities’ of the Membrane Dynamics membrane and structural changes

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 15 modes in bilayers • Incoherent neutron scattering inelastic •NMR • Dielectric spectroscopy Correlated molecular motions might be Local “Humidity Chamber” “Humidity per Si-wafer, mosaicity ~ 0.5° several 1000 bilayers 1000 bilayers several highly oriented Solid supported, membrane stacks with 500 mg „Sandwich-sample“ of deuterated DMPC Stacked Planar Membranes

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 16 Sample Preparation r q z q Scattering from aligned phases Isotropic solution “powder” Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 17 membranes Highly oriented solid supported Si-wafer , 108107 (2004). 93 gneto, Demmel, PRL Salditt, Rheinstädter, Ollinger, Fra Ollinger, Rheinstädter, TAS fluctuations

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 18 Neutron Three-Axes to measure the short wavelength | 0 |Q defines quasi 0 /Q „Brillouin-zone“ π 2 Directed transport? no quantitative theory no quantitative Experiment: Drug Enhancer (and X-rays) access for Low-Q range difficult to fluid gel ) ω , PRL (2004)

et al. s behave “quasi liquid” . Single points in S(Q, Rheinstädter Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 19 argon or liquid helium Short-Range Dispersion Relation on TAS c-atoms of the acyl-chain → Dispersion relation as found in idealDispersion relation liquids as liquid , 048103 (2006). 97 Rheinstädter, Häussler, Salditt, PRL NSE undulations

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 21 Neutron Spin-Echo to measure long wavelength 3 m / J [] ⎞ ⎟ ⎠ d Π ∂ ∂ ⎛ ⎜ ⎝ d − = B ⎫ ⎪ ⎬ ⎪ ⎭ 2 ] u 14 eV 2 . μ 4 ∇ 1 [ = K + ns 2 1 ⎤ ⎥ ⎦ z u ∂ ∂ ⎡ ⎢ ⎣ B ⎧ ⎪ ⎨ ⎪ ⎩ dV ∫ = timescales H Membrane Elasticity m / J [] ⎞ ⎟ ⎠

κ d Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 22 ⎛ ⎜ ⎝ = Bending modulus Compressional modulus K Mesoscopic membrane fluctuations = 100ns = 10ns τ τ S(q,t) 15% = 1ns τ τ λ≅ / = 0.1ns τ Δλ = 0.01ns τ Spin-Echo Technique

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 23 ω good monochromatization good energy resolution Δω / π ) =2 ω Int τ S(q, Average Amplitude 4 point measurement S(q,t)=P(t)=Amp/Aver=0.67 Echo Signals: Characteristics

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 24 ∼1/λ 3 τ IN15 (slow relaxations) IN15 (slow Fits: single exponential 2 τ Spin-Echo Measurements IN11 (fast times)

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 25 1 τ 3 3

B κ η η bulk-elasticity 3 ρ η film Si-wafer Undulations Romanov and Ul’yanov, PRE 66, 061701 (2002) Romanov and PRE Ul’yanov, 66, 061701 Relaxation Dynamics of solid supported bilayers c Softening close to T 2 2 ) )) Undulation Dispersion Relations D D π Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 26 ( Λ s 3 /( ּ 1 π μη T B ( + + 4 4 || || q q 2 || q =10.3 Å =14.5 k d Λ 3 =0.016 Pa / κ η 3 κ η B=1.08 107 J/m3 = ) || q Surface mode ? ( 1 − Bary-Soroker and Diamant, 2006 15, March 871 (2006), 73, Lett., Europhys. Fit to smectic theory hydrodynamic Ribotta, PRL 32, 6, (1974). τ in Membranes 3-axis (IN12) Data+Theory embedded Challenge: Study Dynamics of

47Å

(4,0) (4,0) 0.8

0.45 (3,1)

0.45

(5,0) (2,2)

0.6 0.4

(4,1)

(3,2)

(3,0) (4,0) (3,1)

)

0.35 (3,1) ) (2,2) −1

−1

0.4 (3,0)

(Å

0.4 r (2,2) (Å 600 500 400 (2,1) r

q

q

(2,0) (1,1)

0.2

(1,0) (3,0) 0.35 0

4 3 1 0

0.8 0.6 (meV) 0.4 0.2 10 counts 10 neutron ω Outlook: Dynamics

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 29 Membrane ILL+U Regensburg Karin Schmalzl, DieterKarin Strauch, Acoustic phonon spectrum Sample: Dieter Oesterhelt, MPI Munich MPI Oesterhelt, Dieter Sample: Bacteriorhodopsin in Purple Thanks to •FanniJuranyi, FOCUS, PSI FOCUS, HMI • Röntgenphysik Institut für Salditt, Tim •• Häußler, FRMII, TUM Wolfgang • Christoph Ollinger, IRP • Fragneto, Giovanna ILL, D17 Kiefer, • Schmidt, FZ-Jülich, Wolfgang IN12 • Demmel, ILL, IN3 Franz • Hiess, ILL, IN8 Arno •FanniJuranyi, Seydel, IN10/IN16 Tilo ••Klaus Klaus Habicht, HMI •• Hauss, HMI Thomas • Dieter Richter, FZ-Jülich • Dieter Strauch, Regensburg • Karin Schmalzl, FZ-Jülich Dieter Oesterhelt, MPI for Biochemistry •• Institut Laue-Langevin DFG, Grant SA 772/8-2

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 30 BS TAS || q s μ <1 τ Relaxation Propagation <5000Å & 1ps< ║ DMPC –d54 /q π 2Å<2 Oscillation& Relaxation

Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 31 Collective Excitations in model membranes , 061908 (2005). , 011907 (2007). , 048103 (2006). , 108107 (2004). 71 75 97 93 NSE Rheinstädter et al., PRE Rheinstädter et al., PRE Rheinstädter et al., PRL Rheinstädter et al., PRL The ‘Neutron Window’ Maikel Rheinstädter, NCNR Summer School, Gaithersburg, June 2007 25-29, 32