Document 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.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.