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Optical emission of a conjugated polyelectrolyte: calcium-induced conformational changes in calmodulin and calmodulin-calcineurin interactions
Linköping University, Department of Physics, Chemistry and Biology. Linköping University, The Institute of Technology.ORCID iD: 0000-0002-5582-140X
Linköping University, Department of Physics, Chemistry and Biology. Linköping University, The Institute of Technology.
2004 (English)In: Macromolecules, ISSN 0024-9297, E-ISSN 1520-5835, Vol. 37, no 24, 9109-9113 p.Article in journal (Refereed) Published
Abstract [en]

Electronic polymers in aqueous media offer bioelectronic detection of biomolecular processes. Here we report fluorometric detection of calcium-induced conformational changes in calmodulin based on noncovalent assembly of calmodulin to a water-soluble zwitterionic polythiophene derivative. Assembly with calmodulin will induce a planar geometry and aggregation of the polymer chains, detected as a decrease of the intensity and a red shift of the fluorescence. Upon addition of Ca2+ the intensity of the emitted light is increased and blue-shifted. The geometrical alteration of the polymer chains can further be utilized for recording of the binding of calcineurin to the calcium-activated POWT−calmodulin complex. This novel methodology, using a conformation-sensitive probe, allows fluorometric detection of conformational changes in biomolecules and protein−protein interactions without any covalent modifications of the biomolecules. The rapid and selective method is based on noncovalent interactions between a zwitterionic polythiophene derivative and the biomolecule of interest. This offers a novel way to create microarrays without using covalent attachment of the receptor or labeling of the analyte.

Place, publisher, year, edition, pages
2004. Vol. 37, no 24, 9109-9113 p.
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:liu:diva-45577DOI: 10.1021/ma048605tOAI: oai:DiVA.org:liu-45577DiVA: diva2:266473
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2017-12-13
In thesis
1. Conjugated polyelectrolytes: conformation sensitive optical probes for the recording of biological processes
Open this publication in new window or tab >>Conjugated polyelectrolytes: conformation sensitive optical probes for the recording of biological processes
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The physical properties of conjugated polymers can be utilized for a wide range of biosensors. For instance, the conformational flexibility fouud in conjugated polyelectrolytes, allows direct connection between the geometry of chains and the resulting electronic structure and optical processes, since the extension of the π-conjugated system is distorted by conformational changes of the polyelectrolyte backbone. The biosensors presented in this thesis are utilizing conformational changes of conjugated polyelectrolytes for the detection of biomolecular processes, such as biospecific interactions and conformational changes of biomolecules. The methodology have been used for the detection of DNA-hybridization, single nucleotide polymorphism (SNP) in DNA, conformational alterations of synthetic peptides, conformational alterations of Calmodulin and binding of Ca2+-activated Calmodulin (CaM) to Calcineurin, and amyloid fibril formation of amyloidogenic proteins.

The method is based on non-covalent assembly of a conjugated polyelectrolyte and a biomolecule of interest. Upon exposure to a second biomolecule recognizing the first biomolecule or a conformational change of the first biomolecule, a conformational alteration of the polyelectrolyte backbone and a change in the electronic properties of the polyelectrolyte occurs, and these alterations can be detected by a change of the absorption or the fluorescence from the polyelectrolyte. Hence, conjugated polyelectrolytes can be used as novel conformation sensitive optical probes for the detection of several biological processes. The biomolecular interaction or the conformational changes of the biomolecule are reflected as an alteration of the geometry and the electronic structure of the bouud polyelectrolyte chains and are detected by absorption and emission. The present mechanism may be used for detection of a variety biomolecular processes, and the simplicity and the diversity of this methodology make it suitable for making inexpensive protein- and DNA-chips for rapid detection of biomolecular recognition.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2005. 61 p.
Series
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 961
National Category
Natural Sciences
Identifiers
urn:nbn:se:liu:diva-30214 (URN)15709 (Local ID)91-85457-01-9 (ISBN)15709 (Archive number)15709 (OAI)
Public defence
2005-09-15, Planck, Fysikhuset, Linköpings universitet, Linköping, 13:15 (Swedish)
Opponent
Available from: 2009-10-09 Created: 2009-10-09 Last updated: 2014-04-08

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Nilsson, PeterInganäs, Olle

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