Date of Award
1-1-2009
Document Type
Masters Thesis
Degree Name
M.S.
Organizational Unit
Biological Sciences
First Advisor
Robert M. Dores, Ph.D.
Second Advisor
Cynthia V. Fukami
Third Advisor
Joseph Angleson
Fourth Advisor
Scott Barbee
Keywords
ACTH, Adrenocorticotropic hormone, Activation, CHO-K1, MC2R, Melanocortin 2 receptor, Melanocortin, Silurana Tropicalis
Abstract
Melanocortin receptor ligand selectivity has been a question not easily answered. The inability to functionally express melanocortin 2 receptor (MC2R) has inhibited the study of why MC2R is only stimulated by ACTH, a melanocortin hormone. With the recent discovery of the MC2R accessory protein (MRAP), creating a heterologous system is now feasible. Using a general cell line like CHO-K1 cells, which do not express endogenous MCRs, we were able to create a heterologous expression system and test the selectivity of MC2R using analog variants of ACTH(1-24). Our results indicate an amino acid requirement in the C-terminal portion of ACTH(1-24) for activation, which supports the 2-step method of activation hypothesized for MC2R. This site, the tetra basic cleavage site, when altered does not stimulate cAMP production and does not compete with ACTH(1-24) for binding. We also demonstrate the potential for a non-mammalian MC2R system in cloning full length Silurana tropicalis MC2R and completed localization studies with this system with MRAP using CHO-K1 cells.
Publication Statement
Copyright is held by the author. User is responsible for all copyright compliance.
Rights Holder
Kristopher D. Veo
Provenance
Received from ProQuest
File Format
application/pdf
Language
en
File Size
78 p.
Recommended Citation
Veo, Kristopher D., "Amino Acid Residues Implicated in the Interaction of Melanocortin Ligands and Their Receptors: A Study of MC2R Selectivity" (2009). Electronic Theses and Dissertations. 945.
https://digitalcommons.du.edu/etd/945
Copyright date
2009
Discipline
Molecular Biology