Research Mentor(s)
Anthony-Cahill, Spencer J.
Description
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.
Document Type
Event
Start Date
18-5-2017 9:00 AM
End Date
18-5-2017 12:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
Hemoglobin--Reactivity; Hemoglobin--Stability; Oxygen--Physiological transport; Protein binding
Type
Image
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
Included in
Structural Studies to Understand the Effects of N108K T-State Stabilizing Mutation in Circularly Permuted Hemoglobin
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.