ArticleAnalytical chemistry2013
Measurement of protein tyrosine phosphatase activity in single cells by capillary electrophoresis.
Article in Analytical chemistry, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 30 citations in OpenAlex.
- Cross-Species Applications of Peptide Substrate Reporters to Quantitative Measurements of Kinase Activity.ACS measurement science au · 2024Article
- Photoactivatable Reporter to Perform Multiplexed and Temporally Controlled Measurements of Kinase and Protease Activity in Single Cells.Analytical chemistry · 2021Article
- Advances of Single-Cell Protein Analysis.Cells · 2020Review
- Single-cell analysis targeting the proteome.Nature reviews. Chemistry · 2020Review
- Clinical and pharmaceutical applications of affinity ligands in capillary electrophoresis: A review.Journal of pharmaceutical and biomedical analysis · 2020Review
- Label-Free Assay of Protein Tyrosine Phosphatase Activity in Single Cells.Analytical chemistry · 2019Article
- Design of an automated capillary electrophoresis platform for single-cell analysis.Methods in enzymology · 2019Review
- Advances in enzyme substrate analysis with capillary electrophoresis.Methods (San Diego, Calif.) · 2018Review
- Design and Application of Sensors for Chemical Cytometry.ACS chemical biology · 2018Review
- Development of β-Hairpin Peptides for the Measurement of SCF-Family E3 Ligase Activity in Vitro via Ornithine Ubiquitination.ACS omega · 2017Article
- Ultrasensitive proteomic quantitation of cellular signaling by digitized nanoparticle-protein counting.Scientific reports · 2016Article
- Rational Design of a Dephosphorylation-Resistant Reporter Enables Single-Cell Measurement of Tyrosine Kinase Activity.ACS chemical biology · 2016Article
- Identification of a p53-based portable degron based on the MDM2-p53 binding region.The Analyst · 2016Article
- In vitro methods to study bubble-cell interactions: Fundamentals and therapeutic applications.Biomicrofluidics · 2016Review
- Separation of peptide fragments of a protein kinase C substrate fused to a β-hairpin by capillary electrophoresis.Analytical and bioanalytical chemistry · 2015Article
- Pronase E-Based Generation of Fluorescent Peptide Fragments: Tracking Intracellular Peptide Fate in Single Cells.Analytical chemistry · 2015Article
- Inhibition of the lymphoid tyrosine phosphatase: the effect of zinc(II) ions and chelating ligand fragments on enzymatic activity.Bioorganic & medicinal chemistry letters · 2014Article
- Cellular biochemistry methods for investigating protein tyrosine phosphatases.Antioxidants & redox signaling · 2014Review
- Response of single leukemic cells to peptidase inhibitor therapy across time and dose using a microfluidic device.Integrative biology : quantitative biosciences from nano to macro · 2014Article
- Ex vivo chemical cytometric analysis of protein tyrosine phosphatase activity in single human airway epithelial cells.Analytical chemistry · 2014Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
A fluorescent peptide substrate was used to measure dephosphorylation by protein tyrosine phosphatases (PTP) in cell lysates and single cells and to investigate the effect of environmental toxins on PTP activity in these systems. Dephosphorylation of the substrate by PTPN1 and PTPN2 obeyed Michaelis-Menten kinetics, with KM values of 770 ± 250 and 290 ± 54 nM, respectively. Dose-response curves and IC50 values were determined for the inhibition of these two enzymes by the environmental toxins Zn(2+) and 1,2-naphthoquinone, as well as pervanadate. In A431 cell lysates, the reporter was a poor substrate for peptidases (degradation rate of 100 ± 8.2 fmol min(-1) mg(-1)) but an excellent substrate for phosphatases (dephosphorylation rate of 1.4 ± 0.3 nmol min(-1) mg(-1)). Zn(2+), 1,2-naphthoquinone, and pervanadate inhibited dephosphorylation of the reporter in cell lysates with IC50 values of 470 nM, 35 μM, and 100 nM, respectively. Dephosphorylation of the reporter, following loading into living single cells, occurred at rates of at least 2 pmol min(-1) mg(-1). When single cells were exposed to 1,2-naphthoquinone (50 μM), Zn(2+) (100 μM), and pervandate (1 mM), dephosphorylation was inhibited with median values and first and third quartile values of 41 (Q1 = 0%, Q3 = 96%), 50 (Q1 = 46%, Q3 = 74%), and 53% (Q1 = 36%, Q3 = 77%), respectively, demonstrating both the impact of these toxic exposures on cell signaling and the heterogeneity of response between cells. This approach will provide a valuable tool for the study of PTP dynamics, particularly in small, heterogeneous populations such as human biopsy specimens.
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