Event Title

Generation of Polymeric Recombinant Hemoglobin Using DBCO-modified Hemoglobin

Research Mentor(s)

Spencer Anthony-Cahil

Description

Oligomerized hemoglobin (Hb) molecules have been shown to decrease previously observed deleterious effects from treatment with cell-free hemoglobin1. To create these poly-Hb constructs, a method has developed which employs the site specific ligation reaction of the sortase A enzyme from S. aureus. Hb mutants generated in our lab (αcpβ) have been further modified by adding the sortase recognition sequence, LPXTG, to the C-terminus of the α-subunit (s-αcpβ). Sortase-mediated ligation (SML) will be employed to ligate Hb subunits to a peptide linker that contains a strained cyclooctyne (DBCO), which in turn we plan to react with scaffolding molecules that display azides. We propose to control the final oligomerization state of the poly-Hb by the structure of the scaffold, which will have a defined number of reactive azides that can form covalent bonds with the DBCO-modified Hbs.

Document Type

Event

Start Date

15-5-2019 9:00 AM

End Date

15-5-2019 5:00 PM

Location

Carver Gym (Bellingham, Wash.)

Department

Chemistry

Genre/Form

student projects, posters

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 document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.

Language

English

Format

application/pdf

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May 15th, 9:00 AM May 15th, 5:00 PM

Generation of Polymeric Recombinant Hemoglobin Using DBCO-modified Hemoglobin

Carver Gym (Bellingham, Wash.)

Oligomerized hemoglobin (Hb) molecules have been shown to decrease previously observed deleterious effects from treatment with cell-free hemoglobin1. To create these poly-Hb constructs, a method has developed which employs the site specific ligation reaction of the sortase A enzyme from S. aureus. Hb mutants generated in our lab (αcpβ) have been further modified by adding the sortase recognition sequence, LPXTG, to the C-terminus of the α-subunit (s-αcpβ). Sortase-mediated ligation (SML) will be employed to ligate Hb subunits to a peptide linker that contains a strained cyclooctyne (DBCO), which in turn we plan to react with scaffolding molecules that display azides. We propose to control the final oligomerization state of the poly-Hb by the structure of the scaffold, which will have a defined number of reactive azides that can form covalent bonds with the DBCO-modified Hbs.