Methods for protein labeling based on acyl carrier protein

Details Methods for protein labeling based on acyl carrier protein Methods for protein labeling based on acyl carrier protein

G - Physics

01

N

G01N 33/532 (2006.01) C07K 14/705 (2006.01) C07K 19/00 (2006.01) G01N 33/533 (2006.01) G01N 33/534 (2006.01) G01N 33/535 (2006.01)

Patent

CA 2526579

A method for labeling acyl carrier protein (ACP) fusion proteins with a wide variety of different labels is disclosed. The method relies on the transfer of a label from a coenzyme A type substrate to an ACP fusion protein using a holo- acyl carrier protein synthase (ACPS) or a homologue thereof. The method allows detecting and manipulating the fusion protein, both in vitro and in vivo, by attaching molecules to the fusion proteins that introduce a new physical or chemical property to the fusion protein. Examples of such labels are, among others, spectroscopic probes or reporter molecules, affinity tags, molecules generating reactive radicals, cross-linkers, ligands mediating protein-protein interactions or molecules suitable for the immobilization of the fusion protein.

L'invention porte sur un procédé de marquage de protéines de fusion à protéines transporteuses d'acyles (ACP) en ayant recours à une large palette de différents marqueurs. Le procédé repose sur le transfert d'un marqueur à partir d'un substrat de type coenzyme A vers une protéine de fusion ACP au moyen d'une protéine synthase transporteuse d'acyles (ACPS) ou un de ses homologues. Ce procédé permet de détecter et/ou de manipuler la protéine de fusion, à la fois in vitro et in vivo, en attachant des molécules aux protéines de fusion qui introduisent une nouvelle propriété physique ou chimique dans la protéine de fusion. Parmi les exemples de ces marqueurs on trouve, entre autres, des sondes spectroscopiques ou des molécules rapporteuses, des marqueurs d'affinité, des molécules générant des radicaux réactifs, des agents de réticulation, des ligands qui induisent des interactions protéines-protéines ou des molécules qui servent à immobiliser la protéine de fusion.

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