121st General Meeting of the KCS

Type Poster Presentation
Area Analytical Chemistry
Room No. Event Hall
Time 4월 19일 (목요일) 11:00~12:30
Code ANAL.P-301
Subject Efficient purification processes for expressed human melanocortin-4 receptor
Authors Minseon Kim, soyeon Jo1, Ji Sun Kim2, YONGAE KIM2,*
Department of chemistry, Hankuk University of Foreign Studies, Korea
1chemistry, Hankuk University of Foreign Studies, Korea
2Department of Chemistry, Hankuk University of Foreign Studies, Korea
Abstract Since the human transmembrane proteins (hTMPs) perform various biological functions such as ion channels, signal transduction, and intercellular communication, it is worth identifying the structure of the hTMPs. Membrane proteins are also related to various human diseases, therefore it is attracting attention as a purpose of new medicine development. Human melanocortin-4 receptor (hMC4R) is one of hTMPs related to eating disorder and obesity because it is main controller of body weight and energy homeostasis. Multiple different mutations in hMC4R are inheritable reasons of severe obesity. Especially, it has been called that D90N mutation, which is 90th amino acid, aspartic acid located second transmembrane domain (TM2) was substituted to asparagine. It was found in the patients with early onset obesity. We consider that functional loss of mutant hMC4R-TM2(m- hMC4R-TM2) is caused by structure change of wild type hMC4R-TM2 (wt-hMC4R-TM2). To study functional difference of m-hMC4R-TM2 and wt-hMC4R-TM2 in the progression of early onset obesity, we have confirmed three-dimensional structure of two different peptides. It is important to obtain high yield and high purity of expressed proteins for investigate the structure of wt/m-hMC4R-TM2. The FPLC using sodium dodecyl sulfate (SDS) as detergent due to hydrophobicity of target proteins was applied to separate the target transmembrane proteins from impurities. The transmembrane protein obtained with higher yield and purity after removing SDS was applied to various biophysical techniques like MS, CD, solution NMR spectroscopy, and solid-state NMR spectroscopy.
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