Melanocortin 4 receptors interact with antimicrobial frog peptide analogues

Ernest, U.; Jo, Eun Bae; Choi, Gyu; Piao, Long Zhu; Shin, Jaekyoon; Seo, Min‐Duk; Kang, Su-Jin; Lee, Bong-Jin; Kim, Kang Ho; Kim, Jae Bum; Kim, Su‐Il

doi:10.1016/j.bbrc.2006.03.082

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…Fig. 3 Correlation between the determined K d values of the investigated peptide-DnaK interactions using the quenching assay and the FP assay; error bars show a confidence interval of 95 % (n03) Table 2 Analysis of significant differences in the quenching effects of analyte peptides compared to the negative control apidaecin 1b (9)(10)(11)(12)(13)(14)(15)(16)(17)(18) in the prescreening test using a one-way ANOVA test (significance level, 0.01 %) and determined K d values (quenching assay) …”

Section: Resultsmentioning

confidence: 99%

“…2c). The corresponding quenching curves of the modified oncocin peptides showed a binding to the protein in the same range as oncocin (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11) and oncocin (12)(13)(14)(15)(16)(17)(18)(19) (Fig. 2c, d).…”

Section: Analysis Of Binding Affinitiesmentioning

confidence: 92%

“…As a consequence, medium-to high-throughput methods are needed to determine the binding affinity of proline-rich AMP to DnaK. So far, this screening is mostly based on fluorescence polarization (FP) [9][10][11][12] and surface plasmon resonance [13,14]. FP involves fluorophore labeling of a small ligand (e.g., oncocin) followed by the recording of a binding curve based on the FP measurement with increasing concentrations of the binding partner (e.g., DnaK).…”

Section: Introductionmentioning

confidence: 99%

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Novel peptide–protein assay for identification of antimicrobial peptides by fluorescence quenching

et al. 2012

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The specific interaction of peptides with proteins is often a key factor which determines biological activities. The determination of K(d) values of such interactions is commonly performed with fluorescence polarization. However, fluorescence polarization assays are prone to false-positive results due to the potential for non-specific interactions and only afford very low signal-to-background ratios. Here, we present as an alternative a fluorescence resonance energy transfer based quenching assay to measure peptide-protein interactions in solution. In a test setup where antimicrobial peptides were tested for their affinity towards the protein DnaK, the assay provided high specificity and good reproducibility and correlated with the results obtained by fluorescence polarization methods. Furthermore, we established a fast prescreening method which will allow a highly efficient screening of peptide libraries by reducing the amount of sample by 98% compared to conventional fluorescence polarization assays.

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Section: Resultsmentioning

confidence: 99%

Section: Analysis Of Binding Affinitiesmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

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Novel peptide–protein assay for identification of antimicrobial peptides by fluorescence quenching

et al. 2012

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“…It was suggested that studying nature-derived peptides confer advantages for developing GPCR ligands as drug discover effort [ 189 ]. Full agonists were identified from frog peptides that can bind to human MC4R with good affinities, although EC 50 s are in the tens of micromolar range [ 190 ].…”

Section: Nonclassical Ligandsmentioning

confidence: 99%

Ligands for Melanocortin Receptors: Beyond Melanocyte-Stimulating Hormones and Adrenocorticotropin

Yuan

Tao

2022

Biomolecules

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The discovery of melanocortins in 1916 has resulted in more than 100 years of research focused on these peptides. Extensive studies have elucidated well-established functions of melanocortins mediated by cell surface receptors, including MSHR (melanocyte-stimulating hormone receptor) and ACTHR (adrenocorticotropin receptor). Subsequently, three additional melanocortin receptors (MCRs) were identified. Among these five MCRs, MC3R and MC4R are expressed primarily in the central nervous system, and are therefore referred to as the neural MCRs. Since the central melanocortin system plays important roles in regulating energy homeostasis, targeting neural MCRs is emerging as a therapeutic approach for treating metabolic conditions such as obesity and cachexia. Early efforts modifying endogenous ligands resulted in the development of many potent and selective ligands. This review focuses on the ligands for neural MCRs, including classical ligands (MSH and agouti-related peptide), nonclassical ligands (lipocalin 2, β-defensin, small molecules, and pharmacoperones), and clinically approved ligands (ACTH, setmelanotide, bremelanotide, and several repurposed drugs).

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Homogeneous Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs in Budded Baculoviruses: The Case of Cy3B-NDP-α-MSH Binding to MC4 Receptors

Veikšina

Kopanchuk

Mazina

et al. 2015

Methods in Molecular Biology

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Despite the availability of numerous conceptually different approaches for the characterization of ligand-receptor interactions, there remains a great requirement for complementary methods that are suitable for kinetic studies, especially for the characterization of membrane protein systems and G protein-coupled receptors (GPCRs) in particular. One of the potential approaches that inherently fits well for this purpose is fluorescence anisotropy (FA), a method that allows continuous monitoring of ligand binding processes and characterization of ligand binding dynamics. However, significant changes in FA signal of fluorescently labeled ligands can be detected only if the ratio of bound to free fluorescent ligand portions is altered, which means that receptor and ligand concentrations have to be comparable. As most of the GPCRs are normally present at relatively low concentrations in native tissues and conventional receptor preparations from overexpressed systems often generate high background levels due to significant autofluorescence, receptor preparations with sufficiently high receptor concentrations have become a critical requirement for successful FA assay performance. We propose that budded baculoviruses that display GPCRs on their surfaces can be used as a receptor source in FA assays. Here, we describe the experimental setup of this homogeneous budded baculovirus/FA-based assay system for investigation of receptor-ligand interactions and a novel strategy for FA kinetic data analysis that is taking into account the effect of nonspecific interactions and the depletion of the fluorescent ligand during the binding reaction. The developed budded baculovirus/FA-based assay system brings the experimental data to a level that could solve complex models of ligand-receptor interactions and become a valuable tool for the screening of pharmacologically active compounds. Melanocortin 4 (MC4) receptors and the fluorescent ligand Cy3B-NDP-α-MSH were used as the model system.

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Melanocortin 4 receptors interact with antimicrobial frog peptide analogues

Cited by 7 publications

References 31 publications

Novel peptide–protein assay for identification of antimicrobial peptides by fluorescence quenching

Novel peptide–protein assay for identification of antimicrobial peptides by fluorescence quenching

Ligands for Melanocortin Receptors: Beyond Melanocyte-Stimulating Hormones and Adrenocorticotropin

Homogeneous Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs in Budded Baculoviruses: The Case of Cy3B-NDP-α-MSH Binding to MC4 Receptors

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