Introduction to spin label electron paramagnetic resonance spectroscopy of proteins
作者： Michelle Melanson , Abha Sood , Fanni Török , Marianna Török
DOI: 10.1002/BMB.20677
关键词:
摘要: An undergraduate laboratory exercise is described to demonstrate the biochemical applications of electron paramagnetic resonance (EPR) spectroscopy. The β93 cysteine residue hemoglobin labeled by covalent binding 3-maleimido-proxyl (5-MSL) and 2,2,5,5-tetramethyl-1-oxyl-3-methyl methanethiosulfonate (MTSL), respectively. excess spin label removed gel-exclusion chromatography. Changes in mobility reporter groups attached protein are monitored EPR While spectral parameters rigidly 5-MSL provide information on rotation whole protein, MTSL bound a more flexible linkage describes local environment interior structure. Students can study known crystal structure comparison results they obtain analyzing spectra. Overall, introduces them techniques such as labeling, gel filtration, spectroscopy, well familiarizes with online Protein Data Bank research resource PyMOL software visualization tool.
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