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

Direct link
A Microfluidic Platform for Real-Time Detection and Quantification of Protein-Ligand Interactions
Univ Cambridge, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England..
Univ Coll Dublin, Sch Biomol & Biomed Sci, Dublin 2, Ireland..
Lund Univ, Dept Biochem & Struct Biol, Lund, Sweden..
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology. Lund Univ, Dept Biochem & Struct Biol, Lund, Sweden..
Show others and affiliations
2016 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 110, no 9, 1957-1966 p.Article in journal (Refereed) PublishedText
Abstract [en]

The key steps in cellular signaling and regulatory pathways rely on reversible noncovalent protein-ligand binding, yet the equilibrium parameters for such events remain challenging to characterize and quantify in solution. Here, we demonstrate a microfluidic platform for the detection of protein-ligand interactions with an assay time on the second timescale and without the requirement for immobilization or the presence of a highly viscous matrix. Using this approach, we obtain absolute values for the electrophoretic mobilities characterizing solvated proteins and demonstrate quantitative comparison of results obtained under different solution conditions. We apply this strategy to characterize the interaction between calmodulin and creatine kinase, which we identify as a novel calmodulin target. Moreover, we explore the differential calcium ion dependence of calmodulin ligand-binding affinities, a system at the focal point of calcium-mediated cellular signaling pathways. We further explore the effect of calmodulin on creatine kinase activity and show that it is increased by the interaction between the two proteins. These findings demonstrate the potential of quantitative microfluidic techniques to characterize binding equilibria between biomolecules under native solution conditions.

Place, publisher, year, edition, pages
2016. Vol. 110, no 9, 1957-1966 p.
National Category
URN: urn:nbn:se:uu:diva-297787DOI: 10.1016/j.bpj.2016.03.038ISI: 000375896400006PubMedID: 27166804OAI: oai:DiVA.org:uu-297787DiVA: diva2:943602
EU, European Research CouncilSwedish Research Council
Available from: 2016-06-28 Created: 2016-06-28 Last updated: 2016-06-28Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Persson, Jonas
By organisation
Clinical Immunology
In the same journal
Biophysical Journal

Search outside of DiVA

GoogleGoogle Scholar
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

Altmetric score

Total: 16 hits
ReferencesLink to record
Permanent link

Direct link