120th General Meeting of the KCS

Type Oral Presentation
Area Oral Presentation of Young Analytical Chemists II
Room No. Room C308+C309
Time FRI 10:05-:
Code ANAL2.O-23
Subject The structural characterization of disease related human transmembrane proteins using the NMR spectroscopy
Authors Seongjin Cho, Ji Sun Kim, YONGAE KIM*
Department of Chemistry, Hankuk University of Foreign Studies, Korea
Abstract Human transmembrane proteins (hTMPs) play essential roles in cellular metabolism, participating in processes such as ion transport, nutrient uptake, signal transduction, and intercellular communication. However, aggregation or misfolding of hTMPs due to unwanted mutations of amino acid sequences or errors in the folding process leads to an increase in various human diseases. Therefore, we aim to create new treatments that can inhibit or eliminate these changes. In this research, we tried to identify the structure of human amyloid-β (hAβ) and human melanocortin-4 receptor (hMC4R). The hAβ transmembrane protein shows that the non-fibrillar hAβs form the Ca2+-permeable ion channel in the cell membrane and these channels can disrupt the normal cellular calcium homeostasis. It plays an important role in pathogenesis of dementia and Alzheimer disease. The human melanocortin-4 receptor (hMC4R) has been highlighted because it is central regulator of body weight. Heterozygous mutations related in genetic cause of severe obesity. We succeed to produce the transmembrane domain of the hAβ and second transmembrane domain of the wild-type hMC4R and mutant hMC4R. In cases of the wt/m-TM2 purification, we used sodium dodecyl sulfate (SDS) because of their hydrophobicity. Since SDS binds so strongly to protein, it interferes with biophysical techniques used for characterization of wt/m TM2. Thus, we developed the removal methods of SDS to obtain high quality of purification for wt/m-TM2. Highly purified proteins were applied to several analysis techniques like PAGE, CD, MASS, solution and solid-state NMR spectroscopy. In addition, we could get 1H-15N 1D, 2D SAMMY and SAMPI spectra using home-built solid-state NMR probe.
E-mail sungjin9111@naver.com