Mitochondrial cytochrome c is a heme containing enzyme with a pivotal role in the electron transport train and programmed cell death. The heme is covalently attached to a Cys-X-X-Cys-His motif in cytochrome c by a heme maturation enzyme, holocytochrome c synthase (HCCS). Previous work has taken the iron atom in the heme and chemically substituted it with cobalt to yield a hydrogen-evolution catalyst, but the production of this catalyst is inefficient. This project focuses on studying the possibility of directly expressingsion cobalt-substituted cytochrome c by first creating an expression vector for HCCS using a pGEX backbone with a GST tag via DNA recombination using seamless litigation cloning extract, or SLiCE. Once the vector is verified, variants of the protein will be expressed and used to synthesize cobalt-cytochrome c from apocytochrome c and cobalt-protoporphyrin IX. Currently, the sequence of the pGEX vector with the iLUV gene has been verified, PCR primers have been designed, and PCR has been optimized leading to the successful removal of the iLUV gene in preparation to add the HCCS gene. The HCCS gene, originally in a pBTR vector, has been amplified. DNA purification followed by SLiCE cloning and mutagenesis will yield the expression vectors needed to test the ability of HCCS to directly express metal-substituted cytochrome c.

Creation of an expression vector for holocytochrome c synthase tosubstitute cobalt into the porphyrin ring of mitochondrial cytochrome c

Macromolecular Chemistry: The Second Century

[Author Benefits](https://scimeetings.acs.org/?utm_source=pubs_content_marketing&utm_medium=website&utm_campaign=0320_MCF_NPI_Launch_Spring_Homepage&ref=pubs_content_marketing)

[How to use SciMeetings](https://storage.googleapis.com/dl.morressier.com/acs/SciMeetings-ACSSpring2020-HowToGuide_M3020-Final.pdf)

[Frequently Asked Questions](https://storage.googleapis.com/dl.morressier.com/acs/SciMtng_ACSNMSp2020_FAQ_M3120_Final.pdf)

ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.