| 摘要 |
The invention provides in a first aspect a method for the identification of an ZAP-70 interacting compound, comprising the steps of a) providing a pr otein preparation containing phosphorylated ZAP-70, b) contacting the protei n preparation with aminopyrido-pyrimidine ligand 24 immobilized on a solid s upport under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex, c) incubating the aminopyrido-pyr imidine ligand 24 ~ phosphorylated ZAP-70 complex with a given compound, and d) determining whether the compound is able to separate phosphorylated ZAP- 70 from the immobilized aminopyrido-pyrimidine ligand 24. In a second aspect , the present invention relates to A method for the identification of an ZAP -70 interacting compound, comprising the steps of a) providing a protein pre paration containing phosphorylated ZAP-70, b) contacting the protein prepara tion with ligand aminopyrido-pyrimidine ligand 24 immobilized on a solid sup port and with a given compound under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex, and c) de tecting the aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex formed in step b). hi a third aspect, the invention provides a method for t he identification of an ZAP-70 interacting compound, comprising the steps of : a) providing two aliquots of a protein preparation containing phosphorylat ed ZAP-70, b) contacting one aliquot with aminopyrido-pyrimidine ligand 24 i mmobilized on a solid support under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex, c) contact ing the other aliquot with aminopyrido-pyrimidine ligand 24 immobilized on a solid support and with a given compound under conditions allowing the forma tion of an aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex, and d) determining the amount of aminopyrido- pyrimidine ligand 24 - phosph orylated ZAP-70 complex formed in steps b) and c). In a forth aspect, the in vention relates to a method for the identification of an ZAP-70 interacting compound, comprising the steps of: a) providing two aliquots comprising each at least one cell containing phosphorylated ZAP-70, b) incubating one aliqu ot with a given compound, c) harvesting the cells of each aliquot, d) lysing the cells in order to obtain protein preparations, e) contacting the protei n preparations with aminopyrido-pyrimidine ligand 24 immobilized on a solid support under conditions allowing the formation of an aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex, and f) determining the amount of aminopyrido-pyrimidine ligand 24 - phosphorylated ZAP-70 complex formed in each aliquot in step e).
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