Effects of the Surface Densities of Glycoclusters on the Determination of Their IC50 and Kd Value Determination by Using a Microarray. Issue 16 (6th October 2015)

• Record Type: Journal Article
• Authors: Dupin, Lucie; Zuttion, Francesca; Géhin, Thomas; Meyer, Albert; Phaner‐Goutorbe, Magali; Vasseur, Jean‐Jacques; Souteyrand, Eliane; Morvan, François; Chevolot, Yann
• Is Part Of: Chembiochem. Volume 16:Issue 16(2015)
• Page Start: 2329
• Page End: 2336
• Publication Date: 2015-10-06
• Subjects: Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201500371
• Languages: English
• ISSNs: 1439-4227

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p> <italic>Pseudomonas aeruginosa</italic> (PA) is an opportunistic bacterium involved in 10–30 % of nosocomial diseases. It causes severe lung injury to cystic fibrosis patients, often leading to patient death. PA strains are multidrug resistant, thus making the design of new therapeutics a challenge for public health. One promising therapeutic option is to design glycoclusters that target the virulence factor of PA. <italic>LecA</italic> is a galactose‐specific lectin that might be involved in adhesion and biofilm formation by PA. The DNA‐directed immobilization (DDI) microarray is a powerful tool for screening and understanding of structure–activity relationships between glycoclusters and lectins. High‐throughput and multiplexed analysis of lectin–glycocluster interactions on a DDI microarray allows measurement of IC<sub>50</sub> and dissociation constant (<italic>K</italic><sub>d</sub>) values with minute amounts of material. In order to study the robustness of the DDI microarray in determination of IC<sub>50</sub> and <italic>K</italic><sub>d</sub> values, the impact of glycocluster surface density was investigated. The data obtained show that measured IC<sub>50</sub> values were influenced by glycocluster surface density: as the density of glycoclusters increases, the measured IC<sub>50</sub> values increase too. In contrast, the measured <italic>K</italic><sub>d</sub> values were not affected by glycocluster surface density, provided that the experimental conditions allow interaction between glycocluster and lectin at single‐molecule level (no surface cluster effect).</p> </abstract>