PMID- 36262049
OWN - NLM
STAT- MEDLINE
DCOM- 20221102
LR  - 20221205
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 94
IP  - 43
DP  - 2022 Nov 1
TI  - Super-Resolution Imaging of Autophagy by a Preferred Pair of Self-Labeling 
      Protein Tags and Fluorescent Ligands.
PG  - 15057-15066
LID - 10.1021/acs.analchem.2c03125 [doi]
AB  - Autophagy is a core recycling process for homeostasis, with its dysfunction 
      associated with tumorigenesis and various diseases. Yet, its subtle intracellular 
      details are covered due to the limited resolution of conventional microscopies. 
      The major challenge for modern super-resolution microscopy deployment is the lack 
      of a practical labeling system, which could provide robust fluorescence with 
      fidelity in the context of the dynamic autophagy microenvironment. Herein, a 
      representative autophagy marker LC3 protein is selected to develop two hybrid 
      self-labeling systems with tetramethylrhodamine (TMR) fluorophores through 
      SNAP/Halo-tag technologies. A systematic investigation indicated that the match 
      of the LC3-Halo and TMR ligand remarkably outperforms that of LC3-SNAP, as the 
      former Halo system exhibited more robust single-molecule brightness (440 vs 247), 
      total photon numbers (45600 vs 13500), and dwell time of the initial bright state 
      (0.82 vs 0.40 s) than the latter. With the aid of this desirable Halo system, for 
      the first time, live-cell ferritinophagy is monitored with a spatial resolution 
      of  approximately 50 nm, which disclosed reduced sizes of autophagosomes ( approximately 650 nm, 
      ferritinophagy) than those in nonselective ( approximately 840 nm, mammalian target of 
      rapamycin (mTOR)) and selective autophagy ( approximately 900 nm, mitophagy).
FAU - Man, Huizi
AU  - Man H
AUID- ORCID: 0000-0002-3569-3009
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
FAU - Zhou, Lin
AU  - Zhou L
AUID- ORCID: 0000-0001-6521-5852
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
FAU - Zhu, Guanghao
AU  - Zhu G
AD  - Shanghai Frontiers Science Center for TCM Chemical Biology, Institute of 
      Interdisciplinary Integrative Medicine Research, Shanghai University of 
      Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China.
FAU - Zheng, Ying
AU  - Zheng Y
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
FAU - Ye, Zhiwei
AU  - Ye Z
AUID- ORCID: 0000-0002-4055-3676
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
FAU - Huang, Zhenlong
AU  - Huang Z
AUID- ORCID: 0000-0003-4148-426X
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
FAU - Teng, Xinru
AU  - Teng X
AD  - State Key Laboratory for Chemistry and Molecular Engineering of Medicinal 
      Resources, Collaborative Innovation Center for Guangxi Ethnic Medicine, School of 
      Chemistry and Pharmaceutical Sciences, Guangxi Normal University, 15 Yucai Road, 
      Guilin 541004, China.
FAU - Ai, Chunzhi
AU  - Ai C
AD  - State Key Laboratory for Chemistry and Molecular Engineering of Medicinal 
      Resources, Collaborative Innovation Center for Guangxi Ethnic Medicine, School of 
      Chemistry and Pharmaceutical Sciences, Guangxi Normal University, 15 Yucai Road, 
      Guilin 541004, China.
FAU - Ge, Guangbo
AU  - Ge G
AUID- ORCID: 0000-0002-9670-4349
AD  - Shanghai Frontiers Science Center for TCM Chemical Biology, Institute of 
      Interdisciplinary Integrative Medicine Research, Shanghai University of 
      Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China.
FAU - Xiao, Yi
AU  - Xiao Y
AD  - State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart 
      Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 
      116024, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221019
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Ligands)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Proteins)
SB  - IM
MH  - Ligands
MH  - *Fluorescent Dyes
MH  - *Autophagy
MH  - Mitophagy
MH  - Proteins
EDAT- 2022/10/21 06:00
MHDA- 2022/11/03 06:00
CRDT- 2022/10/20 01:43
PHST- 2022/10/21 06:00 [pubmed]
PHST- 2022/11/03 06:00 [medline]
PHST- 2022/10/20 01:43 [entrez]
AID - 10.1021/acs.analchem.2c03125 [doi]
PST - ppublish
SO  - Anal Chem. 2022 Nov 1;94(43):15057-15066. doi: 10.1021/acs.analchem.2c03125. Epub 
      2022 Oct 19.