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Folding of the Ribosomal protein S6 :The role of sequence connectivity, overlapping foldons, and parallel pathways

by Haglund, Ellinor, PhD

Abstract (Summary)
To investigate how protein folding is affected by sequence connectivity five topological variants of the ribosomal protein S6 were constructed through circular permutation.  In these constructs, the chain connectivity (i.e. the order of secondary-structure elements) is changed without changing the native-state topology.  The effects of the permutations on the folding process were then characterised by ?-value analysis, which estimates the extent of contact formations in the transition-state ensemble.  The results show that the folding nuclei of the wild-type and permutant proteins comprises a common motif of one ?-helix docking against two ?-sheets, i.e. the minimal structure for folding.  However, this motif is recruited in different parts of the S6 structure depending on the permutation, either in the ?1 or ?2 half of the protein.  This minimal structure is not unique for S6 but can also be seen in other proteins.  As an effect of the dual nucleation possibilities, the transition-state changes describe a competition between two parallel pathways, which both include the central ?-stand 1.  This strand constitutes thus a structural overlap between the two competing nuclei.  As similar overlap between competing nuclei is also seen in other proteins, I hypothesise that the coupling of several small nuclei into extended ‘super nuclei’ represents a general principle for propagating folding cooperativity across large structural distances.  Moreover, I demonstrate by NMR analysis that the existence of multiple folding nuclei renders the H/D-exchange kinetics independent of the folding pathway.
Bibliographical Information:

Advisor:

School:Stockholms universitet

School Location:Sweden

Source Type:Doctoral Dissertation

Keywords:NATURAL SCIENCES; Chemistry; Biochemistry; protein folding; protein stability; two-state folding; S6; chevron plot; transition state; parallell pathways; foldon; two-channel landscape; protein engineering; H/D-exchange; biokemi; Biochemistry

ISBN:978-91-7155-939-5

Date of Publication:01/01/2009

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