Folding pathway of guanidine-denatured disulfide-intact wild-type and mutant bovine pancreatic ribonuclease A |
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Authors: | Robert W Dodge John H Laity David M Rothwarf Sakurako Shimotakahara and Harold A Scheraga |
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Institution: | (1) Baker Laboratory of Chemistry, Cornell University, 14853-1301 Ithaca, New York |
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Abstract: | The refolding kinetics of guanidine-denatured disulfide-intact bovine pancreatic ribo-nuclease A (RNase A) and its proline-42-to-alanine mutant (Pro42Ala) have been studied by monitoring tyrosine burial and 2 -cytidine monophosphate (2 CMP) inhibitor binding. The folding rate for wild-type RNase A is faster in the presence of the inhibitor 2 CMP than in its absence, indicating that the transition-state structure in the rate-determining step is stabilized by 2 CMP. The folding rate monitored by 2 CMP binding to the major slow-folding species of Pro42Ala RNase A is faster than the folding rate monitored by tyrosine burial; however, the folding rate monitored by inhibitor binding to the minor slow-folding species is decreased significantly over the folding rate monitored by tyrosine burial, indicating that the major and minor slow-folding species of Pro42Ala fold to the native state with different transition-state conformations in the rate-determining step. |
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Keywords: | Protein folding transition-state structure ligand binding ribonuclease mutant |
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