ProteinFolding-UniversityofNottingham：蛋白质折叠-诺丁汉大学.pptVIP

* * * * * * Protein Folding BiospectroscopyLecture 4 F14PFB David Robinson Protein Folding Introduction Protein Structure Interactions Protein Folding Models Biomolecular Modelling Bioinformatics Protein Folding Protein folding considers the question of how the process of protein folding occurs, i. e. unfolded ? native state. This very challenging problem has been described as the second half of the genetic code, and as the three-dimensional code, as opposed to the one-dimensional code involved in nucleotide/amino acid sequence. Importance: Predict 3D structure from primary sequence Avoid misfolding related to human diseases Design proteins with novel functions Anfinsen Experiment Denaturation of ribonuclease A (4 disulfide bonds), with 8 M Urea containing b-mercaptoethanol, leads to random coil and no activity Anfinsen Experiment After renaturation, the refolded protein has native activity, despite 105 ways to renature the protein. Conclusion: All the information necessary for folding into its native structure is contained in the amino acid sequence of the protein. Anfinsen Experiment Remove b-mercaptoethanol only, oxidation of the sulfhydryl group, then remove urea → scrambled protein, no activity Further addition of trace amounts of b-mercaptoethanol converts the scrambled form into native form. Conclusion: The native form of a protein has the thermodynamic-ally most stable structure. The Levinthal paradox Cyrus Levinthal tried to estimate how long it would take a protein to do a random search of conformational space for the native fold. Many proteins fold in seconds or less: how is this possible? Imagine a 100-residue protein with three possible conformations per residue. Thus, the number of possible folds = 3100 = 5 x 1047. Let us assume that protein can explore new conformations at the same rate that bonds can reorient (1013 structures/second). Thus, the time to explore all of conformational space = 5 x 1047/1013 = 5 x 1034 seconds = 1.6 x 1027 years