Event Title

Clarifying the protein interactions of lycopene elongase and bacterioopsin in H. salinarium

Presenter Information

Adam Lavertu, Colby CollegeFollow

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

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Apr 30th, 9:00 AM Apr 30th, 10:55 AM

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.

http://digitalcommons.colby.edu/clas/2015/program/198