umu.sePublications
Change search
ReferencesLink to record
Permanent link

Direct link
Urea-Dependent Adenylate Kinase Activation following Redistribution of Structural States
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
2016 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 111, no 7, 1385-1395 p.Article in journal (Refereed) Published
Abstract [en]

Proteins are often functionally dependent on conformational changes that allow them to sample structural states that are sparsely populated in the absence of a substrate or binding partner. The distribution of such structural microstates is governed by their relative stability, and the kinetics of their interconversion is governed by the magnitude of associated activation barriers. Here, we have explored the interplay among structure, stability, and function of a selected enzyme, adenylate kinase (Adk), by monitoring changes in its enzymatic activity in response to additions of urea. For this purpose we used a 31P NMR assay that was found useful for heterogeneous sample compositions such as presence of urea. It was found that Adk is activated at low urea concentrations whereas higher urea concentrations unfolds and thereby deactivates the enzyme. From a quantitative analysis of chemical shifts, it was found that urea redistributes preexisting structural microstates, stabilizing a substrate-bound open state at the expense of a substrate-bound closed state. Adk is rate-limited by slow opening of substrate binding domains and the urea-dependent redistribution of structural states is consistent with a model where the increased activity results from an increased rate-constant for domain opening. In addition, we also detected a strong correlation between the catalytic free energy and free energy of substrate (ATP) binding, which is also consistent with the catalytic model for Adk. From a general perspective, it appears that urea can be used to modulate conformational equilibria of folded proteins toward more expanded states for cases where a sizeable difference in solvent-accessible surface area exists between the states involved. This effect complements the action of osmolytes, such as trimethylamine N-oxide, that favor more compact protein states.

Place, publisher, year, edition, pages
2016. Vol. 111, no 7, 1385-1395 p.
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:umu:diva-126432DOI: 10.1016/j.bpj.2016.08.028OAI: oai:DiVA.org:umu-126432DiVA: diva2:1033187
Available from: 2016-10-06 Created: 2016-10-06 Last updated: 2016-10-06

Open Access in DiVA

No full text

Other links

Publisher's full text

Search in DiVA

By author/editor
Rogne, PerWolf-Watz, Magnus
By organisation
Department of Chemistry
In the same journal
Biophysical Journal
Chemical Sciences

Search outside of DiVA

GoogleGoogle Scholar

Altmetric score

Total: 86 hits
ReferencesLink to record
Permanent link

Direct link