Stepwise Assembly of β-Sheet Structure during the Folding of an SH3 Domain Revealed by a Pulsed Hydrogen Exchange Mass Spectrometry Study
| Autor: | Jayant B. Udgaonkar, Nilesh Aghera |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Protein Folding 030102 biochemistry & molecular biology Chemistry Beta sheet Deuterium Exchange Measurement Sequence (biology) Mass spectrometry Biochemistry Mass Spectrometry Protein Structure Secondary SH3 domain src Homology Domains Folding (chemistry) Phosphatidylinositol 3-Kinases 03 medical and health sciences Crystallography 030104 developmental biology Protein structure Protein folding |
| Zdroj: | Biochemistry. 56:3754-3769 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | Dissecting temporally the sequence of secondary structural changes, and determining how these specific changes modulate conformational heterogeneity, remain major goals of protein folding studies. In this study, the folding of the SH3 domain of PI3 kinase has been characterized using pulsed hydrogen exchange mass spectrometry (HX-MS). The folding could be described as a four-state process, U I-vE I-E N, where I-vE and I-E are structurally heterogeneous intermediate ensembles. Compared to U, early intermediate I-VE has a marginally increased level of protection against HX of amides along the entire length of the polypeptide. Sequential assembly into beta-sheet structure has been resolved temporally. Three of the five beta-strands acquire nativelike structure before the rate-limiting step. beta-Strands 2 and 5 acquire nativelike structure in I-vE, while beta-strand 4 does so in I-E. beta-Strand 1 acquires nativelike structure only during the last step of the folding process. Hence, the HX-MS study has resolved the order of assembly of the fi-strands for the formation of the two beta-sheets, which previous studies utilizing Phi-value analysis of several different SH3 domains had been unable to accomplish. Moreover, it is shown that structural heterogeneity decreases in a stepwise manner during the three stages of folding. |
| Databáze: | OpenAIRE |
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