To understand cellular processes at molecular levels, elucidation of protein-protein interactions occurring at a specific location in living cells is required. We have developed a proximity labeling method mediated by the enzyme-mediated activation of radical source (EMARS) reaction, which features a radical formation from labeling reagents by horseradish peroxidase (HRP) set on a molecule of interest (probed molecule). Proximal molecules are covalently labeled with a tag conjugated with the labeling reagent. Here we describe protocols for preparation of a labeling reagent, labeling of neighboring proteins of the probed molecule in living cells, and identification of the labeled proteins.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-4939-9537-0_1 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
The EMARS Reaction for Proximity Labeling.
Methods Mol Biol
March 2020
Department of Biochemistry, Saitama Medical University, Saitama, Japan.
To understand cellular processes at molecular levels, elucidation of protein-protein interactions occurring at a specific location in living cells is required. We have developed a proximity labeling method mediated by the enzyme-mediated activation of radical source (EMARS) reaction, which features a radical formation from labeling reagents by horseradish peroxidase (HRP) set on a molecule of interest (probed molecule). Proximal molecules are covalently labeled with a tag conjugated with the labeling reagent.
View Article and Find Full Text PDFElectronic Medication Administration Records in Long-Term Care Facilities: A Scoping Review.
J Am Geriatr Soc
July 2018
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Objectives: To map the extent, range, and nature of research on the effectiveness, level of use, and perceptions about electronic medication administration records (eMARs) in long-term care facilities (LTCFs) and identify gaps in current knowledge and priority areas for future research.
Design: Scoping review of quantitative and qualitative literature.
Setting: Literature review.
Each GPI-anchored protein species forms a specific lipid raft depending on its GPI attachment signal.
Glycoconj J
October 2015
Center for Innovate and Translational Medicine, Kochi University Medical School, Nankoku, Kochi, 783-8505, Japan.
We previously reported a method, termed enzyme-mediated activation of radical sources (EMARS) for analysis of co-clustered molecules with horseradish peroxidase (HRP) fusion proteins expressed in living cells. This method is featured by radical formation of labeling reagents by HRP. In the current study, we have employed another labeling reagent, fluorescein-conjugated tyramide (FT) instead of the original arylazide compounds.
View Article and Find Full Text PDFExpressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
PLoS One
June 2015
Kochi System Glycobiology Center, Kochi University Medical School, Nankoku, Kochi, Japan; Center for Innovate and Translational Medicine, Kochi University Medical School, Nankoku, Kochi, Japan; Department of Biochemistry, Kochi University Medical School, Nankoku, Kochi, Japan.
Lipid rafts that are enriched in glycosylphosphatidylinositol (GPI)-anchored proteins serve as a platform for important biological events. To elucidate the molecular mechanisms of these events, identification of co-clustering molecules in individual raft domains is required. Here we describe an approach to this issue using the recently developed method termed enzyme-mediated activation of radical source (EMARS), by which molecules in the vicinity within 300 nm from horseradish peroxidase (HRP) set on the probed molecule are labeled.
View Article and Find Full Text PDFIdentification of cell-surface molecular interactions under living conditions by using the enzyme-mediated activation of radical sources (EMARS) method.
Sensors (Basel)
November 2012
Department of Biochemistry, Kochi University Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
Important biological events associated with plasma membranes, such as signal transduction, cell adhesion, and protein trafficking, are mediated through the membrane microdomains. We have developed a novel method termed enzyme-mediated activation of radical sources (EMARS) to identify coclustering molecules on the cell surface under living conditions, which features a radical formation from an aryl azide reagent by horseradish peroxidase (HRP). For identification of molecules labeled by the EMARS reaction, antibody array system and mass spectrometry-based proteomics approaches are available.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!