Method for modifying and identifying functional sites in...

Details Method for modifying and identifying functional sites in... Method for modifying and identifying functional sites in...

G - Physics

01

N

G01N 33/68 (2006.01) G01N 33/533 (2006.01)

Patent

CA 2351667

The invention relates to a method for identifying one or more functional sites in proteins. Said method is characterized by the following steps: a) contacting the target protein with a binding partner (BP tag) linked with a laser-activatable marker (tag), resulting in a complex consisting of target protein and BP tag, b) irradiating the complex consisting of target protein and BP tag with laser light, resulting in the formation of free radicals which selectively modify the bound target protein at the binding site, and c) identifying the selectively modified region of the protein by a combination of protein cleavage and mass spectrometry. The invention further relates to a device for carrying out the inventive method.

L'invention concerne un procédé permettant d'identifier un ou plusieurs sites fonctionnels dans une protéine, qui se caractérise en ce que a) on met en contact la protéine cible avec un partenaire de liaison (BP-Tag) combiné avec un marqueur (Tag) activable par laser, ce qui donne lieu à un complexe composé de la protéine cible et du partenaire de liaison (BP-Tag); b) on soumet le complexe à base de protéine cible et du BP-Tag à l'action de la lumière d'un laser, ce qui permet de produire des radicaux libres qui modifient de manière sélective la protéine cible liée, au point de liaison, et c) on identifie la zone de la protéine modifiée de manière sélective, par une combinaison de dissociation de protéine et de spectrométrie de masse. L'invention concerne en outre un dispositif permettant de mettre ledit procédé en oeuvre.

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