Application of Alanine Scanning to Determination of Amino Acids Essential for Peptide Adsorption at the Solid/Solution Interface and Binding to the Receptor: Surface-Enhanced Raman/Infrared Spectroscopy versus Bioactivity Assays
| Autor: | Helena Domin, Marta Makowska, Grzegorz Burnat, Edyta Proniewicz, Adam Prahl, Izabela Małuch |
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| Rok vydání: | 2021 |
| Předmět: |
Silver
Spectrophotometry Infrared Metal Nanoparticles Infrared spectroscopy Peptide Spectrum Analysis Raman 01 natural sciences Article 03 medical and health sciences symbols.namesake chemistry.chemical_compound Adsorption Drug Discovery Humans 030304 developmental biology chemistry.chemical_classification 0303 health sciences Bombesin Alanine scanning Peptide Fragments 0104 chemical sciences Amino acid Receptors Bombesin 010404 medicinal & biomolecular chemistry HEK293 Cells chemistry Mutagenesis Attenuated total reflection Mutation symbols Biophysics Molecular Medicine Gold Raman spectroscopy Protein Binding |
| Zdroj: | Journal of Medicinal Chemistry |
| ISSN: | 1520-4804 0022-2623 |
| DOI: | 10.1021/acs.jmedchem.1c00397 |
| Popis: | The article describes the application of the alanine-scanning technique used in combination with Raman, surface-enhanced Raman, attenuated total reflection Fourier transform infrared, and surface-enhanced infrared absorption (SEIRA) spectroscopies, which allowed defining the role of individual amino acid residues in the C-terminal 6–14 fragment of the bombesin chain (BN6–14) on the path of its adsorption on the surface of Ag (AgNPs) and Au nanoparticles (AuNPs). A reliable analysis of the SEIRA spectra of these peptides was possible, thanks to a curve fitting of these spectra. By combining alanine-scanning with biological activity studies using cell lines overexpressing bombesin receptors and the intracellular inositol monophosphate assay, it was possible to determine which peptide side chains play a significant role in binding a peptide to membrane-bound G protein-coupled receptors (GPCRs). Based on the analysis of spectral profiles and bioactivity results, conclusions for the specific peptide–metal and peptide–GPCR interactions were drawn and compared. |
| Databáze: | OpenAIRE |
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