NR ATKA
AU Moussa,A.; Cecillon,S.; Coleman,A.W.
TI The Use of para-Sulphonato-calix[n]arenes to Amplify Detection of PrPsc in a Western Blot Assay
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-12
PT Konferenz-Poster
AB
Supramolecular chemistry for their ability to complex a wide range of neutral and charged small organic molecules and also inorganic ions. The biological properties of these molecules have been much slower to come to the fore, however since the 1990s there has been a growing recognition of their potential to interact with proteins.
Given that the structure of the prion contains a known binding site for glycosylaminoglycans, (GAGs) it was of interest to study the interactions of the water soluble para-sulphonato-calix[n]arenes with PrP. Initial Electrospray Mass Spectrometry analysis of the binding of various para-sulphonato-calix[n]arene derivatives with the peptides of PrP responsible for GAG binding showed complexation.
When applied to Western Blot analysis of PrPsc, surprisingly the use of para-sulphonato-calix[n]arenes as co-ligands showed amplification of the detection of bovine brain PrPsc with an antibody specific for the central domain of PrP. In the absence of para-sulphonato-calix[n]arenes the detection limit was at a dilution of 1/FF, in the presence of para-sulphonato-calix[n]arenes the detection limit was lowered to a dilution of 1/VV.
Further studies showed that the amplification of the signal was a factor of both the size of the macrocyclic ring, as might be expected from simple non-specific electrostatic interactions but also was a factor of the substitution at the lower rim of the para-sulphonato-calix[n]arene by either alkoxy carboxylic acid or alkoxy amino groups. This latter finding points to a specific recognition of the para-sulphonato-calix[n]arenes by PrP which leads to a much higher binding by the antibody used in the Western Blot assay.
AD A.Moussa, Agence française pour la Sécurité Sanitaire des Aliments, site de Lyon, France.; Sebastien Cecillon, Anthony W. Coleman, Institut de Biologie et de Chimie des Protéines (CNRS), Lyon, France
SP englisch
PO Deutschland