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Imaging Plasma Membrane Deformations With pTIRFM

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Published on: April 2, 2014

Resolving voltage-dependent structural changes of a membrane photoreceptor by surface-enhanced IR difference

X Jiang¹, E Zaitseva, M Schmidt

¹Department of Biophysical Chemistry (PC III), Bielefeld University, 33615 Bielefeld, Germany.

Proceedings of the National Academy of Sciences of the United States of America

|August 23, 2008

Summary

Surface-enhanced IR difference absorption spectroscopy (SEIDAS) reveals structural changes in membrane proteins. This technique precisely tracks ion transfer events and protein dynamics influenced by membrane potential.

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Area of Science:

Biophysics
Spectroscopy
Membrane protein dynamics

Background:

Membrane proteins, crucial for cellular functions, are challenging to study structurally.
Electrophysiology lacks structural sensitivity for membrane protein analysis.

Purpose of the Study:

To present spectroelectrochemical data on vibrational changes in membrane proteins.
To demonstrate the capability of SEIDAS for resolving structural changes and ion transfer.

Main Methods:

Surface-enhanced IR difference absorption spectroscopy (SEIDAS) applied to a single membrane protein monolayer.
Utilizing vibrational changes perpendicular to the surface to determine angular bond orientations.
Applying negative membrane potentials to observe effects on protein function.

Main Results:

SEIDAS resolved structural changes in sensory rhodopsin II, including the protein backbone and retinal cofactor.
Single ion transfer events were detected and correlated with structural changes.
A negative membrane potential (-0.3 V) selectively halted light-induced proton transfer at D75.
Transmembrane electric fields were found to affect specific proton transfer reactions by raising energy barriers.

Conclusions:

SEIDAS provides unprecedented spatial sensitivity and temporal resolution for studying membrane protein mechanisms.
The technique is effective for investigating voltage-dependent processes in ion channels and transporters.
SEIDAS offers a new approach to elucidate the mechanisms of ion transfer across membranes.