ArticleThe Analyst2012
Metabolism of peptide reporters in cell lysates and single cells.
Article in The Analyst, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
What it found
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
21 citing papers in PubMed, 30 citations in OpenAlex.
- Incorporating a β-hairpin sequence motif to increase intracellular stability of a peptide-based PROTAC.Biochemical engineering journal · 2023Article
- Article
- CPProtectides: Rapid uptake of well-folded β-hairpin peptides with enhanced resistance to intracellular degradation.Peptide science (Hoboken, N.J.) · 2019Article
- "Fix and assay": separating in-cellulo sphingolipid reactions from analytical assay in time and space using an aldehyde-based fixative.The Analyst · 2019Article
- Assays for tyrosine phosphorylation in human cells.Methods in enzymology · 2019Article
- Selection and optimization of enzyme reporters for chemical cytometry.Methods in enzymology · 2019Article
- Development of β-Hairpin Peptides for the Measurement of SCF-Family E3 Ligase Activity in Vitro via Ornithine Ubiquitination.ACS omega · 2017Article
- Development of a protease-resistant reporter to quantify BCR-ABL activity in intact cells.The Analyst · 2016Article
- Rational Design of a Dephosphorylation-Resistant Reporter Enables Single-Cell Measurement of Tyrosine Kinase Activity.ACS chemical biology · 2016Article
- 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
- 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
- Measuring activity in the ubiquitin-proteasome system: from large scale discoveries to single cells analysis.Cell biochemistry and biophysics · 2013Review
- β-Turn sequences promote stability of peptide substrates for kinases within the cytosolic environment.The Analyst · 2013Article
- Measurement of protein tyrosine phosphatase activity in single cells by capillary electrophoresis.Analytical chemistry · 2013Article
- Microfluidic chemical cytometry of peptide degradation in single drug-treated acute myeloid leukemia cells.Analytical chemistry · 2013Article
- Fluorescence lifetime imaging of biosensor peptide phosphorylation in single live cells.Angewandte Chemie (International ed. in English) · 2013Article
- Article
- A multiple reaction monitoring (MRM) method to detect Bcr-Abl kinase activity in CML using a peptide biosensor.PloS one · 2013Article
- Development of a peptidase-resistant substrate for single-cell measurement of protein kinase B activation.Analytical chemistry · 2012Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
The stability of an Abl kinase substrate peptide in a cytosolic lysate and in single cells was characterized. In the cytosolic lysate, the starting peptide was metabolized at an average initial rate of 1.7 ± 0.3 zmol pg(-1) s(-1) with a t(1/2) of 1.3 min. Five different fragments formed over time; however, a dominant cleavage site was identified. Multiple rational design cycles were utilized to develop a lead peptide with a phenylalanine and alanine replaced by an (N-methyl)phenylalanine and isoleucine, respectively, to attain cytosolic peptidase resistance while maintaining Abl substrate efficacy. This lead peptide possessed a 15-fold greater lifetime in the cytosolic lysate while attaining a 7-fold improvement in k(cat) as an Abl kinase substrate compared to the starting peptide. However, when loaded into single cells, the starting peptide and lead peptide possessed nearly identical degradation rates and an altered pattern of fragmentation relative to that in cell lysates. Preferential accumulation of a fragment with cleavage at an Ala-Ala bond in single cells suggested that dissimilar peptidases act on the peptides in the lysate versus single cells. A design strategy for peptide stabilization, analogous to that demonstrated for the lysate, should be effective for stabilization in single cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.