NDLI: Imaging specific newly synthesized proteins within cells by fluorescence resonance energy transfer

Content Provider	Royal Society of Chemistry (RSC)
Author	Liu, Jie Zhang, Xinrong Chen, Hong-Yuan Xu, Jing-Juan Zhang, Sichun Sheng, Linfeng Cai, Lesi
Copyright Year	2017
Abstract	Metabolic azide amino acid labelling followed by the use of bioorthogonal chemistry is an efficient technique for imaging newly synthesized proteins. Recently, AHA-labelling together with the proximity-ligation assay was used to identify newly synthesized proteins of interest (POI) (Tom Dieck et al., Nat. Meth. 2015, 12, 411). Here we build on this study replacing the proximity-ligation assay with FRET to improve the spatial resolution. Herein, we develop a FRET-based strategy for imaging the newly synthesized endogenous POI within cells: a FRET acceptor is installed onto the newly synthesized proteins via click chemistry, and a FRET donor onto the POI via immunocytochemistry. We found that a photobleaching based FRET efficiency imaging mode and a fluorescence lifetime imaging mode showed the distribution of newly synthesized proteins more accurately compared to the direct observation of FRET signals. We demonstrated the capability of this FRET-based imaging method by visualizing several newly synthesized proteins including TDP-43, tubulin and CaMKIIα in different cell lines. This novel analytical imaging method could be used to visualize other specific endogenous proteins of interest in situ.
Starting Page	748
Ending Page	754
Page Count	7
File Format	HTM / HTML PDF
ISSN	20416520
Volume Number	8
Issue Number	1
Journal	Chemical Science
DOI	10.1039/c6sc02610a
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Subscribed
Subject Keyword	Nat Förster resonance energy transfer Immunocytochemistry CAMK2A Fluorescence microscope Tom Dieck Tubulin Carboxylic acid Click chemistry Azide Bioorthogonal chemistry TARDBP Photobleaching
Content Type	Text
Resource Type	Article
Subject	Chemistry

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Clickable fluorescent dyes for multimodal bioorthogonal imaging

Applications of Azide-Based Bioorthogonal Click Chemistry in Glycobiology

Review Applications of Azide-Based Bioorthogonal Click Chemistry (2013)

Single-molecule photophysics of oxazines on DNA and its application in a FRET switch

Fluorescence resonance energy transfer-based hybridization chain reaction for in situ visualization of tumor-related mRNA

An upconversion nanocomposite for fluorescence resonance energy transfer based cholesterol-sensing in human serum

Bioorthogonal Chemistries for Labeling Living Systems

Förster Resonance Energy Transfer

Multiparameter fluorescence image spectroscopy to study molecular interactions

Imaging specific newly synthesized proteins within cells by fluorescence resonance energy transfer

Similar Documents

Clickable fluorescent dyes for multimodal bioorthogonal imaging

Applications of Azide-Based Bioorthogonal Click Chemistry in Glycobiology

Review Applications of Azide-Based Bioorthogonal Click Chemistry (2013)

Single-molecule photophysics of oxazines on DNA and its application in a FRET switch

Fluorescence resonance energy transfer-based hybridization chain reaction for in situ visualization of tumor-related mRNA

An upconversion nanocomposite for fluorescence resonance energy transfer based cholesterol-sensing in human serum

Bioorthogonal Chemistries for Labeling Living Systems

Förster Resonance Energy Transfer

Multiparameter fluorescence image spectroscopy to study molecular interactions

Imaging specific newly synthesized proteins within cells by fluorescence resonance energy transfer