uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
A structurally heterogeneous transition state underlies coupled binding and folding of disordered proteins
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
Stockholm Univ, Dept Biochem & Biophys, Stockholm, Sweden.
Sapienza Univ Roma, Ist Pasteur, Fdn Cenci Bolognetti, Rome, Italy; Sapienza Univ Roma, Ist Biol Patol & Mol, CNR, Dipartimento Sci Biochim A Rossi Fanelli, Rome, Italy.
Show others and affiliations
2019 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 294, no 4, p. 1230-1239Article in journal (Refereed) Published
Abstract [en]

Many intrinsically disordered proteins (IDPs) attain a well-defined structure in a coupled folding and binding reaction with another protein. Such reactions may involve early to late formation of different native structural regions along the reaction pathway. To obtain insights into the transition state for a coupled binding and folding reaction, we performed restrained molecular dynamics simulations using previously determined experimental binding phi(b) values of the interaction between two IDP domains: the activation domain from the p160 transcriptional co-activator for thyroid hormone and retinoid receptors (ACTR) and the nuclear co-activator binding domain (NCBD) of CREB-binding protein, each forming three well-defined alpha-helices upon binding. These simulations revealed that both proteins are largely disordered in the transition state for complex formation, except for two helices, one from each domain, that display a native-like structure. The overall transition state structure was extended and largely dynamic with many weakly populated contacts. To test the transition state model, we combined site-directed mutagenesis with kinetic experiments, yielding results consistent with overall diffuse interactions and formation of native intramolecular interactions in the third NCBD helix during the binding reaction. Our findings support the view that the transition state and, by inference, any encounter complex in coupled binding and folding reactions are structurally heterogeneous and largely independent of specific interactions. Furthermore, experimental phi(b) values and Bronsted plots suggested that the transition state is globally robust with respect to most mutations but can display more native-like features for some highly destabilizing mutations, possibly because of Hammond behavior or ground-state effects.

Place, publisher, year, edition, pages
2019. Vol. 294, no 4, p. 1230-1239
Keywords [en]
intrinsically disordered protein, pre-steady-state kinetics, protein folding, protein-protein interaction, molecular dynamics
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:uu:diva-377701DOI: 10.1074/jbc.RA118.005854ISI: 000457879500014PubMedID: 30514761OAI: oai:DiVA.org:uu-377701DiVA, id: diva2:1291485
Funder
Swedish Research Council, 2016-04965Available from: 2019-02-25 Created: 2019-02-25 Last updated: 2019-02-25Bibliographically approved

Open Access in DiVA

fulltext(1865 kB)68 downloads
File information
File name FULLTEXT01.pdfFile size 1865 kBChecksum SHA-512
1bb28f7f48b0e0eb095c4b949305dac589e64c87621fb59b837389ae46e399e969ea2eaa5b1d7ac2f2c4f6c1f59fc7a95a1dd4ff0584c124e682fae0d3c9fb6c
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Authority records BETA

Karlsson, ElinAndersson, EvaJemth, Per

Search in DiVA

By author/editor
Karlsson, ElinAndersson, EvaJemth, PerCamilloni, Carlo
By organisation
Department of Medical Biochemistry and Microbiology
In the same journal
Journal of Biological Chemistry
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

Search outside of DiVA

GoogleGoogle Scholar
Total: 68 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 162 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf