Covalent Self-Labeling of Tagged Proteins with Chemical Fluorescent Dyes in BY-2 Cells and Arabidopsis Seedlings.
Arabidopsis
/ cytology
Arabidopsis Proteins
/ chemistry
Endocytosis
Fluorescent Dyes
/ chemistry
Microtubule-Associated Proteins
/ genetics
O(6)-Methylguanine-DNA Methyltransferase
/ chemistry
Plant Cells
/ chemistry
Plants, Genetically Modified
Rhodamines
/ chemistry
Seedlings
Time-Lapse Imaging
Tubulin
/ chemistry
Journal
The Plant cell
ISSN: 1532-298X
Titre abrégé: Plant Cell
Pays: England
ID NLM: 9208688
Informations de publication
Date de publication:
10 2020
10 2020
Historique:
received:
09
06
2020
revised:
16
07
2020
accepted:
31
07
2020
pubmed:
9
8
2020
medline:
7
5
2021
entrez:
9
8
2020
Statut:
ppublish
Résumé
Synthetic chemical fluorescent dyes promise to be useful for many applications in biology. Covalent, targeted labeling, such as with a SNAP-tag, uses synthetic dyes to label specific proteins in vivo for studying processes such as endocytosis or for imaging via super-resolution microscopy. Despite its potential, such chemical tagging has not been used effectively in plants. A major drawback has been the limited knowledge regarding cell wall and membrane permeability of the available synthetic dyes. Of 31 synthetic dyes tested here, 23 were taken up into BY-2 cells, while eight were not. This creates sets of dyes that can serve to measure endocytosis. Three of the dyes that were able to enter the cells, SNAP-tag ligands of diethylaminocoumarin, tetramethylrhodamine, and silicon-rhodamine 647, were used to SNAP-tag α-tubulin. Successful tagging was verified by live cell imaging and visualization of microtubule arrays in interphase and during mitosis in Arabidopsis (
Identifiants
pubmed: 32763980
pii: tpc.20.00439
doi: 10.1105/tpc.20.00439
pmc: PMC7534461
doi:
Substances chimiques
Arabidopsis Proteins
0
Fluorescent Dyes
0
Microtubule-Associated Proteins
0
PIN2 protein, Arabidopsis
0
Rhodamines
0
SPIRAL2 protein, Arabidopsis
0
Tubulin
0
tetramethylrhodamine
62669-72-1
O(6)-Methylguanine-DNA Methyltransferase
EC 2.1.1.63
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3081-3094Commentaires et corrections
Type : CommentIn
Informations de copyright
© 2020 American Society of Plant Biologists. All rights reserved.
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