Title

Modifying Muscarinic Signaling through Mutation of the Neurotransmitter Receptor M2's G-Protein

Presenter Information

Hannah Frye

Department

Biological Sciences

Major

Chemistry, Biochemistry emphasis

Research Advisor

Aronstam, Robert

Advisor's Department

Biological Sciences

Funding Source

cDNA Resource Center, Department of Biological Sciences

Abstract

Muscarinic receptors are a class of G-protein-coupled receptors which recognize Acetylcholine to activate three main signaling systems. The path that the signal follows is determined by the G-protein with which the receptor communicates. This study in particular investigates two major receptor subtypes: M2 and M3. Chinese Hamster Ovary (CHO) cells stably transfected with human M2 muscarinic receptor DNA signal through the G-protein G Gai, which normally inhibits formation of cyclic adenosine monophosphate (cAMP) and does not elicit any measurable calcium response. However, CHO cells transfected with M3 signal through the G-protein Gaq, which produces a strong influx of extracellular calcium. To easily study the M2 pathway, we have developed and tested a chimeric protein of these two G-proteins, named Gaqi. This increases the affinity of the altered protein to the M2 receptor. When M2 CHO cells are transfected with this altered DNA, the receptor follows a typical M3-Gaq pathway, producing a measurable calcium response.

Biography

Hannah is a second year student at Missouri University of Science and Technology studying Chemistry with a Biochemistry emphasis. She has been working in Dr. Robert Aronstam's Neurobiology laboratory for one year as an undergraduate researcher. On campus, Hannah is a Resident Assistant in Thomas Jefferson Hall, Public Relations Officer of the Missouri S& T International Genetically Engineered Machines Team, Secretary of the S& T student branch of American Chemical Society, and a brother of the Beta Delta chapter of Alpha Chi Sigma. This upcoming summer of 2013, she will be working for Cargill Com Milling North America as a Quality Management Chemist Intern. She would like to thank Dr. Robert Aronstam, Adam Martin, Vanessa Kaighin and HsiuJen Wang for their invaluable support and assistance throughout her research.

Research Category

Sciences

Presentation Type

Oral Presentation

Document Type

Presentation

Award

Sciences poster session, Third place

Location

Upper Atrium/Hallway

Presentation Date

03 Apr 2013, 9:00 am - 11:45 am

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Apr 3rd, 9:00 AM Apr 3rd, 11:45 AM

Modifying Muscarinic Signaling through Mutation of the Neurotransmitter Receptor M2's G-Protein

Upper Atrium/Hallway

Muscarinic receptors are a class of G-protein-coupled receptors which recognize Acetylcholine to activate three main signaling systems. The path that the signal follows is determined by the G-protein with which the receptor communicates. This study in particular investigates two major receptor subtypes: M2 and M3. Chinese Hamster Ovary (CHO) cells stably transfected with human M2 muscarinic receptor DNA signal through the G-protein G Gai, which normally inhibits formation of cyclic adenosine monophosphate (cAMP) and does not elicit any measurable calcium response. However, CHO cells transfected with M3 signal through the G-protein Gaq, which produces a strong influx of extracellular calcium. To easily study the M2 pathway, we have developed and tested a chimeric protein of these two G-proteins, named Gaqi. This increases the affinity of the altered protein to the M2 receptor. When M2 CHO cells are transfected with this altered DNA, the receptor follows a typical M3-Gaq pathway, producing a measurable calcium response.