University of Kentucky
Generating Tools for Studying the Processes Underlying Energy Conserving Biochemical Reactions
Grade Level at Time of Presentation
Junior
Major
Biochemistry
Minor
Physics
Institution 23-24
University of Kentucky
KY House District #
3
KY Senate District #
12
Faculty Advisor/ Mentor
Anne-Frances Miller, PhD
Department
Department of Chemistry
Abstract
The electron transport flavoprotein (ETF) found in mitochondria mediates transfer of electrons in respiration. However anaerobic bacteria such as Acidaminococcus fermentans possess a different type of ETF, which is able to execute electron transfer bifurcation. A. fermentans ETF (AfeETF) couples two electron transfer reactions, producing a more energetic product than the starting material. This is a newly recognized mechanism that naturally conserves energy at the level of electron flow. Thus, the ETF protein is a valuable source of insight for creating novel materials and devices to capture and store energy. The knowledge of how ETF conformational (open or closed) changes are coupled to the reactivity of its cofactor flavin is essential for understanding how to make the electron flow of such reactions more energy efficient. This project aims to test small extensions to the ETF proteins, that will act as steric doorstops, to hold the protein in its closed or open conformation. These extensions are small polypeptide chains that will be attached to the N terminus or C terminus subunit of the AfeETF. Failure of the new constructs to fold properly will be observed via diminished yield or protein fractionation into the insoluble phase after expression, via SDS PAGE. Integrity will be assessed via the stoichiometry of bound flavin and their optical spectra. Being able to hold the conformation in the open or closed state will in turn allow us to control ETF conformation while testing other reaction variables for energy efficiency. Thus, this project will provide key biochemical knowledge to harness biological reactions for sustainable energy solutions.
Generating Tools for Studying the Processes Underlying Energy Conserving Biochemical Reactions
The electron transport flavoprotein (ETF) found in mitochondria mediates transfer of electrons in respiration. However anaerobic bacteria such as Acidaminococcus fermentans possess a different type of ETF, which is able to execute electron transfer bifurcation. A. fermentans ETF (AfeETF) couples two electron transfer reactions, producing a more energetic product than the starting material. This is a newly recognized mechanism that naturally conserves energy at the level of electron flow. Thus, the ETF protein is a valuable source of insight for creating novel materials and devices to capture and store energy. The knowledge of how ETF conformational (open or closed) changes are coupled to the reactivity of its cofactor flavin is essential for understanding how to make the electron flow of such reactions more energy efficient. This project aims to test small extensions to the ETF proteins, that will act as steric doorstops, to hold the protein in its closed or open conformation. These extensions are small polypeptide chains that will be attached to the N terminus or C terminus subunit of the AfeETF. Failure of the new constructs to fold properly will be observed via diminished yield or protein fractionation into the insoluble phase after expression, via SDS PAGE. Integrity will be assessed via the stoichiometry of bound flavin and their optical spectra. Being able to hold the conformation in the open or closed state will in turn allow us to control ETF conformation while testing other reaction variables for energy efficiency. Thus, this project will provide key biochemical knowledge to harness biological reactions for sustainable energy solutions.