Elucidation of Carbohydrate Molecular Interaction Mechanism of Recombinant and Native ArtinM
| Autor: | Naira C. Pesquero, Paulo Roberto Bueno, Ángel Maquieira, Rosa Puchades, David Giménez-Romero, Isidro S. Monzó |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Models
Molecular PROTEIN ADSORPTION SURFACE KM+ Horseradish peroxidase law.invention symbols.namesake law QUARTZ-CRYSTAL MICROBALANCE BINDING QUIMICA ANALITICA Materials Chemistry Physical and Theoretical Chemistry SPECIFICITY Glycoproteins Binding Sites Chromatography biology Chemistry Lectin Langmuir adsorption model Quartz crystal microbalance Quartz Crystal Microbalance Techniques LECTIN Recombinant Proteins Surfaces Coatings and Films Mannose-Binding Lectins Solvation shell HYDRATION-SHELL Biophysics biology.protein Recombinant DNA symbols Plant Lectins BIOMOLECULAR ADSORPTION ARTOCARPIN Protein adsorption |
| Zdroj: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia instname |
| DOI: | 10.1021/jp403087p |
| Popis: | [EN] The quartz crystal microbalance (QCM) technique has been applied for monitoring the biorecognition of ArtinM lectins at low horseradish peroxidase glycoprotein (HRP) concentrations, using a simple kinetic model based on Langmuir isotherm in previous work.(18) The latter approach was consistent with the data at dilute conditions but it fails to explain the small differences existing in the jArtinM and rArtinM due to ligand binding concentration limit. Here we extend this analysis to differentiate sugar-binding event of recombinant (rArtinM) and native (jArtinM) ArtinM lectins beyond dilute conditions. Equivalently, functionalized quartz crystal microbalance with dissipation monitoring (QCM-D) was used as real-time label-free technique but structural-dependent kinetic features of the interaction were detailed by using combined analysis of mass and dissipation factor variation. The stated kinetic model not only was able to predict the diluted conditions but also allowed to differentiate ArtinM avidities. For instance, it was found that rArtinM avidity is higher than jArtinM avidity whereas their conformational flexibility is lower. Additionally, it was possible to monitor the hydration shell of the binding complex with ArtinM lectins under dynamic conditions. Such information is key in understanding and differentiating protein binding avidity, biological functionality, and kinetics. P.R.B. acknowledges FAPESP (Sao Paulo State Research Fund Agency) for the financial support and all grant projects related to. This research was also funded through project Feder CTQ2010-15943 (CICYT, Spain), GVA ACOMP-2009/650, GVA Prometeo 2010/008, and PAID-06-12 (Universitat Politecnica de Valencia). D.G.-R acknowledges his position to the "Ramon y Cajal" Program (Spanish Ministry of Economy and Competitiveness). We are very grateful to Prof. Dra. Maria Cristina Roque Barreira and her group for providing ArtinM lectins used in this work and to David Gimenez-Pastor for his immense patience. |
| Databáze: | OpenAIRE |
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