APPLICATION OF 1,8-ANS FLUORESCENT PROBE TO IDENTIFY HYDROPHOBIC PATCHES ON THE SURFACE OF NATIVE CA(II) BINDING EF-HAND PROTEINS UPON HEAVY METAL BINDING
Author
Warfel, Olivia Riley
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This item will be available on: 2025-05-01
Abstract
The role that heavy metal ions play in cellular toxicity remains a highly relevant topic of research due to negative health repercussions observed following toxic metal exposure in areas such as Flint, Michigan and Roane County, Tennessee. The goal of the Spuches lab is to understand how calcium, Ca2+, binding EF-hand proteins and peptides interact with toxic metals such as cadmium, Cd2+, and lead, Pb2+, from a fundamental thermodynamic and structural perspective. Specific to this study, human cardiac troponin C (hcTnC), a Ca2+ binding EF-hand protein responsible for cardiac muscle contraction, has been selected as a model system. Previous circular dichroism (CD) data from the Spuches lab indicate that heavy metal ions like Cd2+ can likely induce similar structural changes upon binding to hcTnC as Ca2+. However, both Ca2+ and Cd2+ bind with very different thermodynamic profiles as indicated by isothermal titration calorimetry (ITC). To further investigate the structural changes that occur in the presence of metal ions, the 1,8-ANS fluorescent probe was used to identify hydrophobic patches exposed on the surface of hcTnC upon metal binding. Fluorescence data reveal a significant blue shift upon addition of protein and a minor enhancement in fluorescence intensity. More specifically, the data suggests similar exposure of the hydrophobic surface for the Ca2+ bound, Cd2+ bound, and Pb2+ bound states of hcTnC. Future studies will be conducted to compare this hcTnC data with other known Ca2+ binding EF-hand proteins, such as human skeletal troponin c (hsTnC), in the apo, Ca2+, Cd2+, and Pb2+ bound states to determine if this trend is common amongst all TnC proteins or specific to cardiac muscle.
Date
2023-05-04
Citation:
APA:
Warfel, Olivia Riley.
(May 2023).
APPLICATION OF 1,8-ANS FLUORESCENT PROBE TO IDENTIFY HYDROPHOBIC PATCHES ON THE SURFACE OF NATIVE CA(II) BINDING EF-HAND PROTEINS UPON HEAVY METAL BINDING
(Honors Thesis, East Carolina University). Retrieved from the Scholarship.
(http://hdl.handle.net/10342/13089.)
MLA:
Warfel, Olivia Riley.
APPLICATION OF 1,8-ANS FLUORESCENT PROBE TO IDENTIFY HYDROPHOBIC PATCHES ON THE SURFACE OF NATIVE CA(II) BINDING EF-HAND PROTEINS UPON HEAVY METAL BINDING.
Honors Thesis. East Carolina University,
May 2023. The Scholarship.
http://hdl.handle.net/10342/13089.
May 15, 2024.
Chicago:
Warfel, Olivia Riley,
“APPLICATION OF 1,8-ANS FLUORESCENT PROBE TO IDENTIFY HYDROPHOBIC PATCHES ON THE SURFACE OF NATIVE CA(II) BINDING EF-HAND PROTEINS UPON HEAVY METAL BINDING”
(Honors Thesis., East Carolina University,
May 2023).
AMA:
Warfel, Olivia Riley.
APPLICATION OF 1,8-ANS FLUORESCENT PROBE TO IDENTIFY HYDROPHOBIC PATCHES ON THE SURFACE OF NATIVE CA(II) BINDING EF-HAND PROTEINS UPON HEAVY METAL BINDING
[Honors Thesis]. Greenville, NC: East Carolina University;
May 2023.
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Publisher
East Carolina University