Progress Toward an O2-carrying Therapeutic Based on a Single-chain Hemoglobin Framework
Research Mentor(s)
Anthony-Cahill, Spencer J.; Spiegel, P. Clint
Description
Our research is focused on the production of a hemoglobin based oxygen carrier (HBOC) that can be used as a therapeutic in the event of acute blood loss. In addition, HBOCs have been shown to increase the efficacy of ionizing radiation during cancer treatment. To achieve this goal, the subunits of the cell-free HBOC must not dissociate when transfused. Ideally, the HBOC would have the oxygen carrying characteristics (binding affinity and cooperativity) of whole blood. Our initial design implemented the co-expression of circular permutated β-globin subunits (cpβ) with α-globin subunits (α-cpβ). Subsequent constructs incorporated the covalent linkage of α and β-globin subunits to generate a single-chain α-cpβ heterodimers (sc-α-cpβ). Preliminary experimentation indicated that our permuteins preferred the high affinity R-state conformation over the low affinity T state. This led to the addition of T-state stabilizing point mutations and the covalent linkage between sc-α-cpβ dimers (scHb). Preliminary ligand binding kinetics data for scHb indicate a preference for the T-state. We are currently working on the structural characterization of scHb as well as inclusion of further T-state stabilizing mutations.
Document Type
Event
Start Date
14-5-2015 10:00 AM
End Date
14-5-2015 2:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
Hemoglobin--Reactivity; 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
Progress Toward an O2-carrying Therapeutic Based on a Single-chain Hemoglobin Framework
Our research is focused on the production of a hemoglobin based oxygen carrier (HBOC) that can be used as a therapeutic in the event of acute blood loss. In addition, HBOCs have been shown to increase the efficacy of ionizing radiation during cancer treatment. To achieve this goal, the subunits of the cell-free HBOC must not dissociate when transfused. Ideally, the HBOC would have the oxygen carrying characteristics (binding affinity and cooperativity) of whole blood. Our initial design implemented the co-expression of circular permutated β-globin subunits (cpβ) with α-globin subunits (α-cpβ). Subsequent constructs incorporated the covalent linkage of α and β-globin subunits to generate a single-chain α-cpβ heterodimers (sc-α-cpβ). Preliminary experimentation indicated that our permuteins preferred the high affinity R-state conformation over the low affinity T state. This led to the addition of T-state stabilizing point mutations and the covalent linkage between sc-α-cpβ dimers (scHb). Preliminary ligand binding kinetics data for scHb indicate a preference for the T-state. We are currently working on the structural characterization of scHb as well as inclusion of further T-state stabilizing mutations.