Protein structure determinationSlide 2Slide 3Slide 4X-ray crystallographySlide 6Slide 7Nuclear magnetic resonance spectroscopySlide 9Slide 10Slide 11Slide 12Protein structure determinationTertiary protein structure: protein foldingThree main approaches:[1] experimental determination (X-ray crystallography, NMR)[2] Comparative modeling (based on homology)[3] Ab initio (de novo) prediction (Dr. Ingo Ruczinski at JHSPH)Experimental approaches to protein structure[1] X-ray crystallography-- Used to determine 80% of structures-- Requires high protein concentration-- Requires crystals-- Able to trace amino acid side chains-- Earliest structure solved was myoglobin[2] NMR-- Magnetic field applied to proteins in solution-- Largest structures: 350 amino acids (40 kD)-- Does not require crystallizationSteps in obtaining a protein structure Target selection Obtain, characterize protein Determine, refine, model the structure Deposit in databaseX-ray crystallographyhttp://en.wikipedia.org/wiki/X-ray_diffractionSperm Whale MyoglobinNuclear magnetic resonance
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