Naturally split inteins assemble through a "capture and collapse" mechanism

Neel H. Shah, Ertan Eryilmaz, David Cowburn, Tom W. Muir

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Split inteins are a class of naturally occurring proteins that carry out protein splicing in trans. The chemical mechanism of protein trans-splicing is well-understood and has been exploited to develop several powerful protein engineering technologies. Split intein chemistry is preceded by efficient molecular recognition between two protomers that become intertwined in their bound state. It is currently unclear how this unique topology is achieved upon fragment association. Using biophysical techniques in conjunction with protein engineering methods, including segmental isotopic labeling, we show that one split intein fragment is partly folded, while the other is completely disordered. These polypeptides capture each other through their disordered regions and form an ordered intermediate with native-like structure at their interface. This intermediate then collapses into the canonical intein fold. This mechanism provides insight into the evolutionary constraints on split intein assembly and should enhance the development of split intein-based technologies.

Original languageEnglish (US)
Pages (from-to)18673-18681
Number of pages9
JournalJournal of the American Chemical Society
Volume135
Issue number49
DOIs
StatePublished - Dec 11 2013

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Inteins
Proteins
Protein Splicing
Protein Engineering
Molecular recognition
Trans-Splicing
Polypeptides
Technology
Engineering technology
Labeling
Protein Subunits
Topology
Association reactions
Peptides

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Naturally split inteins assemble through a "capture and collapse" mechanism. / Shah, Neel H.; Eryilmaz, Ertan; Cowburn, David; Muir, Tom W.

In: Journal of the American Chemical Society, Vol. 135, No. 49, 11.12.2013, p. 18673-18681.

Research output: Contribution to journalArticle

Shah, Neel H. ; Eryilmaz, Ertan ; Cowburn, David ; Muir, Tom W. / Naturally split inteins assemble through a "capture and collapse" mechanism. In: Journal of the American Chemical Society. 2013 ; Vol. 135, No. 49. pp. 18673-18681.
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