Degree Type
Honors Capstone Project
Date of Submission
Spring 5-1-2013
Capstone Advisor
Dr. Ivan Korendovych
Honors Reader
Dr. James Hougland
Capstone Major
Biology
Capstone College
Arts and Science
Audio/Visual Component
no
Capstone Prize Winner
no
Won Capstone Funding
yes
Honors Categories
Sciences and Engineering
Subject Categories
Biochemistry | Biochemistry, Biophysics, and Structural Biology
Abstract
Abstract
Metalloenzymes constitute a large subclass of proteins with diverse functions including metal-sensing, signal transduction pathways, and metal transport. In this report, I aim to study the intricate relationship between structure and function as it relates to metalloproteins in the fields of catalysis and bioimaging. For catalysis, I used existing platforms in addition to de novo design to study small scaffolds with varying secondary structure, and their function in catalyzing the hydrolysis of a p-nitrophenol ester. For bioimaging, I used an existing enzyme known as human carbonic anhydrase I, hCA, to develop a bioimaging agent that can be fused to other biological molecules. After subjecting hCA to mutagenesis, I altered its metal binding site to accommodate a rhenium based complex; these results could eventually be applied to Tc99m due to the two metals’ similar reactivity, and used as a bioimaging technique based on technetium’s gamma radiation, which aids in visualization. Similar approaches are under current development but they are based on synthetic peptide probes. The hCA-Re complex is based on a naturally occurring enzyme, which could eliminate any immunogenic response. Finally, all of the described genes were cloned using ligation-independent cloning, LIC, which is an advantageous although uncommonly used cloning method. LIC does not usually require the purchase of a commercial kit, and it does not require DNA ligase, hence the name of the method. In addition, virtually any gene can be cloned using LIC as the gene and vector sequences do not play major roles in LIC design.
Recommended Citation
Zhezherya, Krystyna, "Design of Metalloproteins for Catalysis and Bioimaging" (2013). Renée Crown University Honors Thesis Projects - All. 75.
https://surface.syr.edu/honors_capstone/75
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