Molecular mechanism of voltage-gated Ca2+ channel regulation by membrane PIP2
- Title
- Molecular mechanism of voltage-gated Ca2+ channel regulation by membrane PIP2
- Alternative Title
- 세포막 인지질 PIP2에 의한 전압 개폐 칼슘채널의 활성 조절 메커니즘 규명
- Author(s)
- Park, Cheon-Gyu
- DGIST Authors
- Suh, Byung-Chang ; Park, Cheon-Gyu ; Jang, Deok-Jin
- Advisor
- 서병창
- Co-Advisor(s)
- Jang, Deok-Jin
- Issued Date
- 2020
- Awarded 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
- Degree
- Doctor
- Department
- Brain and Cognitive Sciences
- Publisher
- DGIST
- Related Researcher
-
-
Suh, Byung-Chang
- 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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- Appears in Collections:
- Department of Brain Sciences Theses Ph.D.
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