ATR-FTIR studies in pore forming and membrane induced fusion peptides.

Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, 76100 Israel. Yechiel.Shai@weizmann.ac.il

Biochimica et biophysica acta 2013 Oct

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Infrared (IR) spectroscopy has been shown to be very reliable for the characterization, identification and quantification of structural data. Particularly, the Attenuated Total Reflectance (ATR) technique which became one of the best choices to study the structure and organization of membrane proteins and membrane-bound peptides in biologically relevant membranes. An important advantage of IR spectroscopy is its ability to analyze material under a very wide range of conditions including solids, liquids and gases. This method allows elucidation of component secondary structure elements of a peptide or protein in a global manner, and by using site specific isotope labeling allows determination of specific regions. A few advantages in using ATR-FTIR spectroscopy include; a relatively simple technique, allow the determination of peptide orientation in the membrane, allow the determination of secondary structures of very small peptides, and importantly, the method is sensitive to isotopic labeling on the scale of single amino acids. Many studies were reported on the use of ATR-FTIR spectroscopy in order to study the structure and orientation of membrane bound hydrophobic peptides and proteins. The list includes native and de-novo designed peptides, as well as those derived from trans-membrane domains of various receptors (TMDs). The present review will focus on several examples that demonstrate the potential and the simplicity in using the ATR-FTIR approach to determine secondary structures of proteins and peptides when bound, inserted, and oligomerized within membranes. The list includes (i) a channel forming protein/peptide: the Ca(2+) channel phospholamban, (ii) a cell penetrating peptide, (iii) changes in the structure of a transmembrane domain located within ordered and non-ordered domains, and (iv) isotope edited FTIR to directly assign structure to the membrane associated fusion peptide in context of a Key gp41 Structural Motif. Importantly, a unique advantage of infrared spectroscopy is that it allows a simultaneous study of the structure of lipids and proteins in intact biological membranes without an introduction of foreign perturbing probes. Because of the long IR wavelength, light scattering problems are virtually non-existent. This allows the investigation of highly aggregated materials or large membrane fragments. This article is part of a Special Issue entitled: FTIR in membrane proteins and peptide studies. Copyright © 2012 Elsevier B.V. All rights reserved.