Title

Deciphering the palmitoylation code of ABHD17A for spatiotemporal control of Ras and its role in proliferation suppression in fibrosarcoma

DGIST Authors

Byeol-I Kim ; Byung-Chang Suh ; Jaemin Lee

Advisor

서병창

Co-Advisor(s)

Jaemin Lee

Issued Date

2026

Awarded Date

2026-02-01

Type

Thesis

Description

Palmitoylation, ABHD17A, Depalmitoylase, Ras, Fibrosarcoma

Table Of Contents

Ⅰ. INTRODUCTION 1
ⅠⅠ. MATERIALS AND METHODS 13
2.1 Cell culture and transfection 13
2.2 Plasmids and molecular cloning 13
2.3 Confocal imaging and analysis 16
2.4 Acyl-RAC capture and Acyl-PEG assays 17
2.5 Western blotting 18
2.6 PM-Cytosol separation 18
2.7 MTS assay 19
2.8 Cell photographs and growth 19
2.9 Chemicals 19
2.10 In silico analysis 20
2.11 Statistics analysis 20
IIⅠ. RESULTS 22
3.1 A palmitoylation code for ABHD17A regulating membrane engagement sorting and catalytic positioning 22
3.1.1 Subcellular localization of the ABHD17 family 22
3.1.2 Single N-terminal cysteine codes are insufficient for ABHD17A palmitoylation and membrane association 31
3.1.3 Middle region palmitoylation in ABHD17A is necessary for PM localization and catalytic activity 34
3.1.4 Hydrophobic residues in the N-terminus regulate initial endomembrane engagement prior to palmitoylation 43
3.1.5 Endosomal targeting motif in ABHD17A contributes to PM targeting 49
3.2 Spatiotemporal control of Ras palmitoylation by chemogenetic and optogenetic tools and its impact on signaling and proliferation 56
3.2.1 Chemogenetic recruitment of ABHD17A enables acute depalmitoylation of Ras (ChemoDepalm) 56
3.2.2 Isoform-specific Ras depalmitoylation by ChemoDepalm and analysis of the K-Ras4A hybrid motif 62
3.2.3 ChemoDepalm-mediated depalmitoylation of Ras reduces PI3K/PIP3 signaling and suppresses proliferationin cancer cells 71
3.2.4 Reversible, light-gated depalmitoylation of N-Ras using OptoDepalm 79
ⅠV. DISCUSSION 84
V. REFERENCES 93
VⅠ. SUMMARY IN KOREAN 103

URI

https://scholar.dgist.ac.kr/handle/20.500.11750/59588
http://dgist.dcollection.net/common/orgView/200000946428

DOI

10.22677/THESIS.200000946428

Degree

Doctor

Department

Department of Brain Sciences

Publisher

DGIST

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