title: Evaluating the coupling efficiency of phosphorylated amino acids for SPOT synthesis.
abstract: A high demand of interest concerning binding assays to study the consequences of posttranscriptional phosphorylation may be addressed by peptide array-based methods. A crucial factor for de novo chemical approaches to generate such arrays is the possibility to rationally permutate phosphorylation events along a huge number of sequences. The simple principle behind this advantage is the stepwise synthesis of peptides, which allows the incorporation of either phosphorylated or nonphosphorylated derivates at serine, threonine, and tyrosine positions. In spite of several reported applications of phosphopeptide arrays, there is, to our best knowledge, no reported analysis of the efficiency of the involved techniques. Here, we analyze different coupling conditions to introduce phosphoamino acids in standard SPOT synthesis. Our results clearly indicate that EEDQ is the preferable activator and can also be used in fully automated SPOT synthesis.
authors.affiliation: Institut für Medizinische Immunologie, Charité-Universitätsmedizin Berlin, Hessische Str. 3-4, 10115 Berlin, Germany.
authors.forename: Victor
authors.initials: V
authors.lastname: Tapia
authors.forename: Bernhard
authors.initials: B
authors.lastname: Ay
authors.forename: Julia
authors.initials: J
authors.lastname: Triebus
authors.forename: Eike
authors.initials: E
authors.lastname: Wolter
authors.forename: Prisca
authors.initials: P
authors.lastname: Boisguerin
authors.forename: Rudolf
authors.initials: R
authors.lastname: Volkmer
mesh_terms: D020650:Combinatorial Chemistry Techniques; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry*; D019606:Phosphoamino Acids / Q000737:chemistry*; D010748:Phosphopeptides / Q000138:chemical synthesis / Q000737:chemistry; D040081:Protein Array Analysis
chemical_list: D010455:Peptides; D019606:Phosphoamino Acids; D010748:Phosphopeptides