Development of a Fluorescence Lifetime Based Method to Detect and Analyze Single Molecule Reactions in Solution
Colyer, Ryan Anthony
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Permalink
https://hdl.handle.net/2142/80576
Description
Title
Development of a Fluorescence Lifetime Based Method to Detect and Analyze Single Molecule Reactions in Solution
Author(s)
Colyer, Ryan Anthony
Issue Date
2008
Doctoral Committee Chair(s)
Gratton, Enrico
Department of Study
Physics
Discipline
Physics
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biophysics, General
Language
eng
Abstract
This new hardware was used to observe the conformational dynamics of a calmodulin sample labeled with a FRET pair, and encapsulated within 100nm lipid vesicles. A toolbox of analysis techniques was developed for PTA, and was used to quantitatively describe the transitions and fluctuations of the conformation of calmodulin. Analysis was done in a model-free manner, and also by applying known parameters about the system to extract more specific information. Using the information obtained, a conformational model was developed to describe the dynamic behavior of calmodulin's conformation in terms of its binding with calcium. In addition, measurements conducted in the presence of a peptide derived from Ca2+/calmodulin-dependent protein kinase II were used to examine the properties of calmodulin's conformational dynamics while interacting with its binding targets.
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