Method for characterization of the fine structure of protein...

G - Physics – 01 – N

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G01N 33/53 (2006.01) C12Q 1/37 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2228404

The binding sites of binding proteins and their binding partners are characterized, at the individual amino acid level, by a combination of heavy hydrogen (tritium or deuterium) exchange labeling and sequential degradation (preferably, by carboxypeptidase) and analysis of labeled fragments under slowed exchange conditions. By first labeling the binding partner and labeling the binding site of the protein indirectly (by exchange at the binding surface), the binding site may be differentiated from allosterically protected segments of the binding protein.

Selon l'invention, les sites de liaison de protéines liantes et de leurs partenaires de liaison sont caractérisés, au niveau de l'aminoacide individuel, par combinaison d'un marquage par échange d'hydrogène lourd (tritium ou deutérium) et d'une dégradation séquentielle (de préférence par carboxypeptidase) et analyse de fragments marqués dans des conditions d'échange ralenti. En marquant d'abord le partenaire de liaison et en marquant le site de liaison de la protéine indirectement (par échange à la surface de liaison), il est possible de différencier le site de liaison de segments allostériquement protégés de la protéine liante.

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