Presentation structural proteomics.pptx

its about the techniques to determine the structure of proteins

1. Presented by Manal Nazim Khan Bsbiotec-f19-12 Protein structure and techniques

2. Structural proteomics Structural proteomics is the application of protein chemistry and modern mass spectrometric techniques to problems such as the characterization of protein structures and to determine the 3D configuration. Term

3. Protein structure

4. Techniques for protein structure determination

5. Primary structure determination Edman degradation Edman degradation is the process of purifying protein by sequentially removing one residue at a time from the amino end of a peptide . It was developed by Pher Edmanin 1950 Process

6. Mass spectroscopy Mass spectrometry (MS) is an analytical technique that separates ionized particles such as atoms, molecules, and clusters by using differences in the ratios of their charges to their respective masses (mass/charge; m/z), and can be used to determine the molecular weight of the particles . It includes A n ion source A mass analyzer A detector A computer Limitations Morphological abnormalities in tissue samples observed by traditional microscopes do not provide us with any biochemical information . On the other hand, conventional biochemical techniques generally lose the positional information of the tissues. Due to such limitations today we are using imaging mass spectroscopy (IMS) with several mass analyzer and ionization methods variations.

7. MALDI T he mostly used ionization method is MALDI matrix which is used for ionizing materials. Matrix-Assisted Laser Deposition/Ionization (MALDI) is a sample preparation technique used to prepare an analyte for mass spectrometry. MALDI is a relatively gentle technique, which usually allows molecules to be analyzed without significant fragmentation . Sample preparation TOF mass analyzer

8. ESI Electrospray ionization is another method to ionize peptides and was developed by Fenn and coworkers in 1989. Mechanism

9. Circular dichroism Circular Dichroism , an absorption spectroscopy, uses circularly polarized light to investigate structural aspects of optically active chiral media . Circular Dichroism (CD) is an absorption spectroscopy method based on the differential absorption of left and right circularly polarized light . Terms include: Chiral molecule Electromagnetic waves Linear polarization Circular polarization Ellipse polarization https:// www.youtube.com/watch?v=Fu-aYnRkUgg

10. X-ray crystallography X-ray Crystallography is a scientific method used to determine the arrangement of atoms of a crystalline solid in three dimensional space . Process Crystallization through drop diffusion method Sample preparation X-ray scattering Electron density maps A three dimensional structure of protein

11. NMR Spectroscopy Nuclear magnetic resonance (NMR) spectroscopy is the study of molecules by recording the interaction of radiofrequency ( Rf ) electromagnetic radiations with the nuclei of molecules placed in a strong magnetic field . Nuclei of atoms Sample preparation Principle of NMR Chemical shift

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