Co-Author(s)

Johann Sigurjonsson, P. Clint Spiegel

Research Mentor(s)

Spencer Anthony-Cahill

Affiliated Department

Chemistry

Sort Order

57

Start Date

18-5-2017 9:00 AM

End Date

18-5-2017 12:00 PM

Document Type

Event

Abstract

The primary goal of our project is to develop a functional hemoglobin based oxygen carrier (HBOC) for clinical care. In order to accomplish this, we are working on designing a single chain hemoglobin (scHb) that can serve as the building block of a monodisperse polymeric HBOC. The scHb is formed from novel covalent fusions between the two α chains and two circularly permuted β chains. The initial constructs of non-covalently linked α and cp-β showed decreased stability and low T-state affinity (Asmundson, et. al., 2009). To counter these effects, two point mutations have been included to increase T-state affinity (Tsai et. al in 1999). The first mutation, N108K, shows greater T-state affinity, however the second mutation, V96W, shows decreased T-state affinity. I am conducting X-ray crystallography trials of the βN108K mutant in order to understand the structural basis for this unexpected effect on the T-state stability, and to inform future protein engineering of the HBOC candidates.

Rights

Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this documentation for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.

Language

English

Format

application/pdf

Share

COinS
 
May 18th, 9:00 AM May 18th, 12:00 PM

Structural Studies to Understand the Effects of N108K T-State Stabilizing Mutation in Circularly Permuted Hemoglobin

Chemistry

The primary goal of our project is to develop a functional hemoglobin based oxygen carrier (HBOC) for clinical care. In order to accomplish this, we are working on designing a single chain hemoglobin (scHb) that can serve as the building block of a monodisperse polymeric HBOC. The scHb is formed from novel covalent fusions between the two α chains and two circularly permuted β chains. The initial constructs of non-covalently linked α and cp-β showed decreased stability and low T-state affinity (Asmundson, et. al., 2009). To counter these effects, two point mutations have been included to increase T-state affinity (Tsai et. al in 1999). The first mutation, N108K, shows greater T-state affinity, however the second mutation, V96W, shows decreased T-state affinity. I am conducting X-ray crystallography trials of the βN108K mutant in order to understand the structural basis for this unexpected effect on the T-state stability, and to inform future protein engineering of the HBOC candidates.

 

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.