Fourier transform infrared (FTIR) spectroscopy is a very useful tool for the investigation of protein structure, of the molecular mechanism of protein reactions, as well as of protein folding, unfolding and misfolding. The vibration of the protein backbone amide group, caused by the absorption of radiation in the mid-IR region (400 – 4000 cm−1), is most widely used for protein structural analysis. These vibrations give rise to nine characteristic IR absorption bands (Amides A, B and I–VII). Especially the characteristic frequency, intensity, and width of the Amide I (∼ 1650 cm−1) and Amide II (∼ 1550 cm−1) vibrational bands, which are the most prominent and sensitive bands of the protein backbone, relate directly to specific protein secondary structural components (i.e. α-helix, β-sheet, β-turn). In this way the obtained IR spectrum of a protein sample can be suitably analysed (using algorithms such as second-derivative, deconvolution and curve fitting if necessary) and thus elucidate the protein secondary structure.

National Hellenic Research Foundation (N.H.R.F.)
LOCATION:   Athens, Greece |
RELEVANT STAFF:   Vassilis Gionis

The Instruct-EL hub provides access to different spectroscopic techniques available at TPCI. Specifically, a Fourier Transformed Mid-Infrared spectrometer (Bruker Equinox 55) equipped with an Attenuated Total Reflectance (ATR) diamond accessory (SENS-IR) and spectral range 400 – 6000 cm-1, is dedicated to FTIR measurements of protein samples. It should be noted that the versatility of the ATR accessory allows the direct measurement of protein powders, gels, solutions as well as proteins dried on the ATR surface, especially when the protein quantity and concentration are limited. The corresponding provided services mainly involve the investigation of protein secondary structure, along with conformational changes like unfolding and/or aggregation caused by solution conditions (i.e. pH, ionic strength, temperature) or interactions with other (bio)macromolecules.

Equinox 55 by Bruker
TYPE:   FTIR spectrometer
LOCATION:   Theoretical and Physical Chemistry Institute / Instruct-EL hub
DESCRIPTION:   Studies on protein structure in solutions and solid state
FUNDER:   External funding
The provision of spectroscopic measurements and analyses has been a long-lasting service (since 1997) of the TPCI’s Analytical Spectroscopy Lab. The involved research staff possesses thorough knowledge and hands-on experience of protein structural and conformational studies using FTIR spectroscopy.