Evaluating the coupling efficiency of phosphorylated amino acids for SPOT synthesis.
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.
Tapia
Victor
V
Institut für Medizinische Immunologie, Charité-Universitätsmedizin Berlin, Hessische Str. 3-4, 10115 Berlin, Germany.
Ay
Bernhard
B
Triebus
Julia
J
Wolter
Eike
E
Boisguerin
Prisca
P
Volkmer
Rudolf
R
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
D010455:Peptides; D019606:Phosphoamino Acids; D010748:Phosphopeptides