Can infrared spectroscopy provide information on protein–protein interactions?
- 26 July 2010
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Society Transactions
- Vol. 38 (4), 940-946
- https://doi.org/10.1042/bst0380940
Abstract
For most biophysical techniques, characterization of protein-protein interactions is challenging; this is especially true with methods that rely on a physical phenomenon that is common to both of the interacting proteins. Thus, for example, in IR spectroscopy, the carbonyl vibration (1600-1700 cm(-1)) associated with the amide bonds from both of the interacting proteins will overlap extensively, making the interpretation of spectral changes very complicated. Isotope-edited infrared spectroscopy, where one of the interacting proteins is uniformly labelled with (13)C or (13)C,(15)N has been introduced as a solution to this problem, enabling the study of protein-protein interactions using IR spectroscopy. The large shift of the amide I band (approx. 45 cm(-1) towards lower frequency) upon (13)C labelling of one of the proteins reveals the amide I band of the unlabelled protein, enabling it to be used as a probe for monitoring conformational changes. With site-specific isotopic labelling, structural resolution at the level of individual amino acid residues can be achieved. Furthermore, the ability to record IR spectra of proteins in diverse environments means that isotope-edited IR spectroscopy can be used to structurally characterize difficult systems such as protein-protein complexes bound to membranes or large insoluble peptide/protein aggregates. In the present article, examples of application of isotope-edited IR spectroscopy for studying protein-protein interactions are provided.Keywords
This publication has 7 references indexed in Scilit:
- Two-dimensional IR spectroscopy and isotope labeling defines the pathway of amyloid formation with residue-specific resolutionProceedings of the National Academy of Sciences of the United States of America, 2009
- Solution State NMR Structure and Dynamics of KpOmpA, a 210 Residue Transmembrane Domain Possessing a High Potential for Immunological ApplicationsJournal of Molecular Biology, 2009
- Interaction of a G Protein-coupled Receptor with a G Protein-derived Peptide Induces Structural Changes in both Peptide and Receptor: A Fourier-transform Infrared Study Using Isotopically Labeled PeptidesJournal of Molecular Biology, 2007
- Solution Structure of the Phosphoryl Transfer Complex between the Cytoplasmic A Domain of the Mannitol Transporter IIMannitol and HPr of the Escherichia coliPhosphotransferase SystemPublished by Elsevier BV ,2002
- A New Approach to the Study of Protein−Protein Interaction by FTIR: Complex Formation between Cytochrome P450BM-3 Heme Domain and FMN Reductase DomainBiochemistry, 2002
- Does Fourier-transform infrared spectroscopy provide useful information on protein structures?Trends in Biochemical Sciences, 1992
- Potential of carbon-13 and nitrogen-15 labeling for studying protein-protein interactions using Fourier-transform infrared spectroscopyBiochemistry, 1992