Date of Award
2001
Degree Name
Chemistry
College
College of Science
Type of Degree
M.S.
Document Type
Thesis
First Advisor
Dr. William Price
Second Advisor
Dr. Leslie Frost
Third Advisor
Dr. Michael L. Norton
Fourth Advisor
Dr. Leonard J. Deutsch
Abstract
The chemistry of transport, storage, energy production, and reproduction in physiological systems is controlled by the actions and interactions of proteins. A species full complement of these amazing machines is often referred to as its proteome. The new science of proteomics utilizes a set of powerful and evolving tools to characterize the nature of the proteome, and further to fully elucidate the functional attributes of the proteins and their associations.
A methodology was developed by combining several proteomic techniques, such as electrophoresis, RP-HPLC, and mass spectrometry, to correctly identity proteins. Three known proteins (myoglobin, albumin, and actin) were employed for development of these techniques to achieve optimal results. The meticulously refined methodology was then applied to an unknown protein in Zucker serum to obtain information about the protein’s activity, structure, and possible post-translational modifications.
The protein was identified as apolipoprotein A-I and studied in Zucker serum to analyze its implications for hereditary obesity. Apo A-I was found to be much more abundant in the obese serum than in the lean. Apo A-I is normally found in the phospholipid layer of serum, but this particular protein was found in the aqueous layer of Zucker serum. These results indicate a difference in the protein. Delipidated Zucker serum analyzed by MALDI-TOF revealed several modifications to the protein, each ~200 Da apart. Two-dimensional electrophoresis of the delipidated serum suggested a multitude of differences in protein levels between the lean and obese. This research has only scratched the surface of understanding the role apo A-I plays in hereditary obesity.
Subject(s)
Proteins.
Analytical chemistry.
Biochemistry.
Recommended Citation
Neal, Laura Brumfield, "Methods development for the identification of unknown proteins" (2001). Theses, Dissertations and Capstones. 1748.
https://mds.marshall.edu/etd/1748
