Event Title
Clarifying the protein interactions of lycopene elongase and bacterioopsin in H. salinarium
Location
Parker-Reed, SSWAC
Start Date
30-4-2015 9:00 AM
End Date
30-4-2015 10:55 AM
Project Type
Poster- Restricted to Campus Access
Description
Membrane proteins play vital roles in cellular functions. Further clarification of interactions between proteins will increase our understanding of this fundamental biological process. Integral membrane proteins have proven difficult to study due to their location within the lipid bilayer of the cell membrane. Halobacterium salinarium contains the membrane proteins lycopene elongase (Lye) and bacterioopsin. Previous research demonstrated that bacterioopsin inhibits Lye activity, and we are testing to see whether these two proteins interact. We are constructing new strain of H. salinarium with a modified bacterioopsin gene (bop) to encode a histidine tag on the C terminus. The strain also will include a modified Lye gene with an epitope tag to the end of the protein. We plan to conduct experiments to determine if a formaldehyde cross-linker would bind to bacterioopsin and any other proteins in proximity to it. If Lye and bacterioopsin interact, then they should be close together within the membrane to be cross-linked. Ive also been conducting a phylogenetic analysis to investigate possible co-evolution of the two proteins. Constructing phylogenetic trees based on the protein sequences and then comparing the relationships between trees. To this point, we have generated new plasmids containing the histidine-tagged bop which can be used as a vector to insert a histidine-tagged bop into H.salinarium. We have also developed a control for the immunoblot screening process. The phylogenetic analysis has shown good indications that the two proteins have co-evolved, supporting the existence of interactions.
Sponsoring Department
Colby College. Biology Dept.
CLAS Field of Study
Natural Sciences
Event Website
http://www.colby.edu/clas
ID
1683
Clarifying the protein interactions of lycopene elongase and bacterioopsin in H. salinarium
Parker-Reed, SSWAC
Membrane proteins play vital roles in cellular functions. Further clarification of interactions between proteins will increase our understanding of this fundamental biological process. Integral membrane proteins have proven difficult to study due to their location within the lipid bilayer of the cell membrane. Halobacterium salinarium contains the membrane proteins lycopene elongase (Lye) and bacterioopsin. Previous research demonstrated that bacterioopsin inhibits Lye activity, and we are testing to see whether these two proteins interact. We are constructing new strain of H. salinarium with a modified bacterioopsin gene (bop) to encode a histidine tag on the C terminus. The strain also will include a modified Lye gene with an epitope tag to the end of the protein. We plan to conduct experiments to determine if a formaldehyde cross-linker would bind to bacterioopsin and any other proteins in proximity to it. If Lye and bacterioopsin interact, then they should be close together within the membrane to be cross-linked. Ive also been conducting a phylogenetic analysis to investigate possible co-evolution of the two proteins. Constructing phylogenetic trees based on the protein sequences and then comparing the relationships between trees. To this point, we have generated new plasmids containing the histidine-tagged bop which can be used as a vector to insert a histidine-tagged bop into H.salinarium. We have also developed a control for the immunoblot screening process. The phylogenetic analysis has shown good indications that the two proteins have co-evolved, supporting the existence of interactions.
https://digitalcommons.colby.edu/clas/2015/program/198