A fluorescent light-up probe as an inhibitor of intracellular β-tryptase

Wang, Qi; Shi, Xiuyin; Zhu, Xiaoyang; Ehlers, Martin; Wu, Junchen; Schmuck, Carsten

doi:10.1039/c4cc02208d

Cited by 8 publications

(6 citation statements)

References 21 publications

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“…ASAH1 (N-acylsphingosine amidohydrolase; EC 3.5.1.23) is a lysosomal enzyme which hydrolyses the N-acyl linkage in ceramide to produce sphingosine and free fatty acids. , To determine the inhibitory effect of E2 on acid ceramidase activity in vitro , a fluorescence-based enzymatic activity assay was performed with HUVECs lysate as the source of acid ceramidase. BODIPY C 5 -ceramide was applied as the substrate under acidic conditions (pH 4.5). − It was shown that E2 specifically and competitively inhibited ASAH1 activity since the latter was recovered by increasing the substrate concentration. When the substrate concentration was 20 μM, ASAH1 activity decreased by 61% ( P < 0.0001).…”

Section: Results and Discussionmentioning

confidence: 99%

A Guanidine-Based Synthetic Compound Suppresses Angiogenesis via Inhibition of Acid Ceramidase

et al. 2018

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Angiogenesis generates new blood vessels from pre-existing vessels. Tumors induce the formation of new blood vessels to ensure sufficient oxygen and nutrients for their growth. Normally, angiogenesis is induced by various pro-angiogenesis factors, including vascular endothelial growth factor (VEGF). Inhibition of VEGF is a promising approach to cancer treatment. A guanidine-based synthetic compound, E2, was identified as a potent hit from 68 guanidine-based derivatives by screening for angiogenesis inhibitors showing antiproliferative activity in human umbilical vein endothelial cells (HUVECs). To explore the mode of action of E2, target proteins were investigated using phage display biopanning, and acid ceramidase 1 (ASAH1) was identified as an E2-binding protein. Drug affinity responsive target stability (DARTS) and ASAH1 activity assays revealed the direct binding of E2 to ASAH1. Moreover, siRNA knockdown of ASAH1 demonstrated its role as an angiogenesis factor. Consequently, E2 inhibited chemoinvasion and tube formation of HUVECs in a dose-dependent manner. E2 also potently suppressed neo-vascularization of chorioallantoic membranes in vivo. Collectively, these data suggest that E2 is a novel angiogenesis inhibitor and ASAH1 is proposed to be a new antiangiogenesis target.

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Section: Results and Discussionmentioning

confidence: 99%

A Guanidine-Based Synthetic Compound Suppresses Angiogenesis via Inhibition of Acid Ceramidase

et al. 2018

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“…The reported cationic ligand binds at the rim of anionic residues around the entrance of the central pore, blocks the access to the active sites, and inhibits enzyme activity. Schmuck et al reported a cationic fluorescent peptidic β-tryptase inhibitor (peptide 9, Figure 8) [51]. The peptide is designed by attaching two Lys-Trp-Lys tripeptide units to a central lysine via the C-terminus.…”

Section: β-Tryptasementioning

confidence: 99%

“…Schmuck et al reported a cationic fluorescent peptidic β-tryptase inhibitor (peptide 9 , Fig. 8 ) [ 51 ]. The peptide is designed by attaching two Lys–Trp–Lys tripeptide units to a central lysine via the C-terminus.…”

Section: Reviewmentioning

confidence: 99%

“…Inset: the corresponding titration curve showing the increase of the emission at 400 nm with increasing β-tryptase concentration; B) confocal laser scanning microscopy images of CHMAS cells treated with 10 μM peptide 9 for 30 min. Reproduced from [ 51 ], “A fluorescent light-up probe as an inhibitor of intracellular β-tryptase”, © 2014 Wang et al, licensed under a Creative Commons Attribution 3.0 Unported Licence, https://creativecommons.org/licenses/by/3.0/. Published by The Royal Society of Chemistry.…”

Section: Reviewmentioning

confidence: 99%

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Selected peptide-based fluorescent probes for biological applications

Maity¹

2020

Beilstein J. Org. Chem.

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To understand the molecular interactions, present in living organisms and their environments, chemists are trying to create novel chemical tools. In this regard, peptide-based fluorescence techniques have attracted immense interest. Synthetic peptide-based fluorescent probes are advantageous over protein-based sensors, since they are synthetically accessible, more stable, and can be easily modified in a site-specific manner for selective biological applications. Peptide receptors labeled with environmentally sensitive/FRET fluorophores have allowed direct detection/monitoring of biomolecules in aqueous media and in live cells. In this review, key peptide-based approaches for different biological applications are presented.

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“…In recent years, molecular probes have been broadly applied in the tracking of biological species such as DNA 18 19 20 21 , polysaccharides 22 , cell surface proteins 23 24 , enzymes 25 and even bacteria 26 . In this context, we planned to prepare photochromic-probe-modified antibacterial peptides to investigate their antibacterial activity and potential use in mechanism studies.…”

mentioning

confidence: 99%

Synthesis and Antibacterial Activities of Antibacterial Peptides with a Spiropyran Fluorescence Probe

Chen

Zhu

Yang

et al. 2014

Sci Rep

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In this report, antibacterial peptides1-3 were prepared with a spiropyran fluorescence probe. The probe exhibits a change in fluorescence when isomerized from a colorless spiro-form (spiropyran, Sp) to a colored open-form (merocyanine, Mc) under different chemical environments, which can be used to study the mechanism of antimicrobial activity. Peptides 1-3 exhibit a marked decrease in antimicrobial activity with increasing alkyl chain length. This is likely due to the Sp-Mc isomers in different polar environments forming different aggregate sizes in TBS, as demonstrated by time-dependent dynamic light scattering (DLS). Moreover, peptides 1-3 exhibited low cytotoxicity and hemolytic activity. These probe-modified peptides may provide a novel approach to study the effect of structural changes on antibacterial activity, thus facilitating the design of new antimicrobial agents to combat bacterial infection.

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A fluorescent light-up probe as an inhibitor of intracellular β-tryptase

Abstract: A pyrene-functionalized peptidic inhibitor binds to and inhibits β-tryptase in a non-competitive and reversible manner even in cells. Upon protein binding a fluorescence increase of the two pyrene fluorophores is observed which allows using as a fluorescent light-up probe for this enzyme.

Cited by 8 publications

References 21 publications

A Guanidine-Based Synthetic Compound Suppresses Angiogenesis via Inhibition of Acid Ceramidase

A Guanidine-Based Synthetic Compound Suppresses Angiogenesis via Inhibition of Acid Ceramidase

Selected peptide-based fluorescent probes for biological applications

Synthesis and Antibacterial Activities of Antibacterial Peptides with a Spiropyran Fluorescence Probe

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