Stepwise Assembly of beta-Sheet Structure during the Folding of an SH3 Domain Revealed by a Pulsed Hydrogen Exchange Mass Spectrometry Study

Aghera, Nilesh and Udgaonkar, Jayant B (2017) Stepwise Assembly of beta-Sheet Structure during the Folding of an SH3 Domain Revealed by a Pulsed Hydrogen Exchange Mass Spectrometry Study. In: BIOCHEMISTRY, 56 (29). pp. 3754-3769.

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Abstract

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.

Item Type:	Journal Article
Publication:	BIOCHEMISTRY
Additional Information:	Copy right for this article belongs to the AMER CHEMICAL SOC, 1155 16TH ST, NW, WASHINGTON, DC 20036 USA
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	01 Sep 2017 09:57
Last Modified:	01 Sep 2017 09:57
URI:	http://eprints.iisc.ac.in/id/eprint/57724

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