Protein structure determination. Tertiary protein structure: protein folding Three main approaches:...
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Protein structure determination
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Tertiary protein structure: protein folding
Three 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)
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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 crystallization
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Steps in obtaining a protein structure
Target selection
Obtain, characterize protein
Determine, refine, model the structure
Deposit in database
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X-ray crystallographyhttp://en.wikipedia.org/wiki/X-ray_diffraction
Sperm Whale Myoglobin
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Nuclear magnetic resonance spectroscopy
http://en.wikipedia.org/wiki/Nuclear_magnetic_resonance
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