Degree Type
Honors Capstone Project
Date of Submission
Spring 5-1-2012
Capstone Advisor
Professor Robert Doyle
Honors Reader
Assistant Professor Ivan Korendovych
Capstone Major
Biology
Capstone College
Arts and Science
Audio/Visual Component
no
Capstone Prize Winner
no
Won Capstone Funding
no
Honors Categories
Sciences and Engineering
Subject Categories
Biochemistry | Biochemistry, Biophysics, and Structural Biology | Other Biochemistry, Biophysics, and Structural Biology
Abstract
Although both the α- and β-domains of intrinsic factor (IF) have been previously expressed, the full crystal structure of the protein has yet to be reported. The purpose of this research is to (1) express IF in order to obtain a complete crystal structure and (2) utilize a mutant form of IF in order to orally deliver rotavirus to the ileum.The first goal of this research is to express IF in the yeast Pichia pastoris. The second goal is to express a K159D IF mutant protein. K159 of IF plays a role in salt bridge formation between IF and the CUB6 domain of cubilin, a critical receptor for vitamin B12 enterocyte passage. The third goal is to express a Q201A IF mutant protein. Q201 of intrinsic factor plays a role in the formation of hydrogen bonding between IF and the CUB6 domain of cubilin. We hypothesize the mutants will either prevent binding to cubilin or allow binding, but not allow for receptor-mediated endocytosis. Utilizing the vitamin B12 pathway with the K159D or Q201A mutated IF will allow oral delivery of antigens, such as the rotavirus vaccine, to the ileum where they can trigger an IgA immune response.
Recommended Citation
Cyphers, Soreen, "Recombinant Expression and Purification of Human Intrinsic Factor (IF) and Mutants K159D and Q201A Designed to Interfere with Cubilin Receptor Binding" (2012). Honors Capstone Projects - All. 138.
https://surface.syr.edu/honors_capstone/138
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
