SERB (Grant No: EMR/2014/000491) funded project
Accurate study of hydrogen bonding in proteins demands experimental localization of hydrogen atoms, which is accessible only from ultra-high resolution X-ray and neutron diffraction experiments. Simple crystal structure analysis would only provide the geometrical features, whereas charge density analysis using combined high-resolution X-ray and neutron data would allow extracting precise structural information in terms of its detailed electron density distributions. Here we plan to use this charge density analysis tool in a novel way to (i) construct the charge density model of protein structures, determined using ultra-high resolution X-ray and neutron diffraction data collected on crystals grown from freshly synthesized proteins, (ii) perform quantitative analysis N-H…O=Cestimate their (iii) gain detailed and predictive understanding of how local atomic density distributions and electrostatic surface modifications direct the preferences in protein folding.
- Expression, purification, perdeuteration (exchanging the H-atoms with its isotope D-atoms), and crystallization of selected protein systems.
- Ultra-high resolution protein structure determination using both X-ray and neutron diffraction data.
- Detail investigation of electron density distributions obtained from advanced protein charge density analysis.
- Quantitative characterization of N-H…O=C hydrogen bonds, the key building blocks of both a-helix and β-sheet structures.
Project funded by SERB (Grant No: EMR/2014/000491)