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Molecular mechanism of voltage-gated Ca2+ channel regulation by membrane PIP2

Title
Molecular mechanism of voltage-gated Ca2+ channel regulation by membrane PIP2
Translated Title
세포막 인지질 PIP2에 의한 전압 개폐 칼슘채널의 활성 조절 메커니즘 규명
Authors
Park, Cheon-Gyu
DGIST Authors
Park, Cheon-Gyu; Jang, Deok-Jin; Suh, Byung-Chang
Advisor(s)
서병창
Co-Advisor(s)
Jang, Deok-Jin
Issue Date
2020
Available Date
2020-06-15
Degree Date
2020-02
Type
Thesis
Description
Voltage-gated calcium (CaV) channel, CaV β subunit, phosphatidylinositol 4,5-bisphosphate (PIP2), molecular gating mechanism
Table Of Contents
Abstract ……………………………………………………………………………………………………………………… i List of contents …………………………………………………………………………………………………………… iii List of tables ………………………………………………………………………………………………………………… v List of figures ………………………………………………………………………………………………………………… v 1. INTRODUCTION ………………………………………………………………………………………………………… 1 2. MATERIALS AND METHODS ………………………………………………………………………………………… 8 2-1. cDNA …………………………………………………………………………………………………………………… 8 2-2. Molecular cloning …………………………………………………………………………………………………… 8 2-3. Cell culture and transfection ……………………………………………………………………………………… 9 2-4. Electrophysiology …………………………………………………………………………………………………… 10 2-5. Confocal imaging …………………………………………………………………………………………………… 10 2-6. Preparation of liposomes ………………………………………………………………………………………… 11 2-7. Assay for peptide-liposome binding ……………………………………………………………………………… 11 2-8. Curve fitting ………………………………………………………………………………………………………… 11 2-9. Förster resonance energy transfer (FRET) ……………………………………………………………………… 12 2-10. Statistical analysis …………………………………………………………………………………………………… 12 3. RESULTS …………………………………………………………………………………………………………………… 15 3-1. The HOOK region of β2 subunit regulates inactivation kinetics and PIP2 sensitivity of CaV2.2 channels ………………………………………………………………………………………………………… 15 3-2. Charged amino acids in the HOOK region mainly determine gating properties of CaV2.2 channels ………………………………………………………………………………………………………… 27 3-3. The short form of the HOOK region in β2c-short subunit shows similar regulatory effects on PIP2 sensitivity and inactivation of CaV2.2 channels ……………………………………………………………… 35 3-4. The B-domain of the HOOK region tethers β subunit to plasma membrane via electrostatic and hydrophobic interactions …………………………………………………………………………………………… 38 3-5. Distance between plasma membrane and CaV β subunit is critically important for modulating inactivation kinetics and PIP2 sensitivity of CaV2.2 channels …………………………………………………… 46 3-6. Conformational shift of I-II loop mainly determines gating properties of CaV2.2 channels ………… 53 3-7. Polybasic residues at the C-terminal end of I-II loop mainly determine PIP2 and muscarinic modulation of CaV2.2 channels ………………………………………………………………………………………… 66 4. DISCUSSION……………………………………………………………………………………………………………… 72 5. REFERENCES …………………………………………………………………………………………………………… 84 6. SUMMARY IN KOREAN ……………………………………………………………………………………………… 92
URI
http://dgist.dcollection.net/common/orgView/200000281554
http://hdl.handle.net/20.500.11750/11920
DOI
10.22677/Theses.200000281554
Degree
Doctor
Department
Brain and Cognitive Sciences
University
DGIST
Related Researcher
  • Author Suh, Byung-Chang Laboratory of Brain Signal and Synapse Research
  • Research Interests Molecular mechanisms of epilepsy and sensory pain transmission; Signaling mechanism of ion channel regulation and membrane excitability; 분자전기생리; 간질 및 통증의 분자적 기전 연구
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Collection:
Department of Brain and Cognitive SciencesThesesPh.D.


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