NOVEL METHOD FOR PRODUCING PROTEIN HAVING A CARBON-CARBON BOND-BASED SITE-SPECIFIC MODIFICATION

• Main Authors: PARK, Hee-Sung, LEE, Hee-Yoon, YANG, Aerin, HA, Sura
• Format: Patent
• Language: English; French; Korean
• Published: 08.03.2018
• Subjects: CHEMISTRY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; METALLURGY; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; TESTING

The present invention is related to a method for producing a target protein having a particular site which has been modified, more specifically to a novel method for producing a target protein having a carbon-carbon bond-based site-specific modification, the method comprising the steps of: (a) marking the site to be modified with a substance having an effector by adding amino acid to the particular site in the target protein; (b) activating the marker at the site; and (c) harvesting the modified protein by adhering a post-translational modification (PTM) moiety on the activated marker. The method for producing a target protein having a site-specific modification according to the present invention not only adds the desired modification at the desired site of the target protein, the target protein to which a modification has been added also exhibits the same effect as the target protein present inside the cell, and thus the present invention could be useful in researching target protein function, screening novel drugs, and the like. La présente invention concerne un procédé de production d'une protéine cible ayant un site particulier qui a été modifié, plus spécifiquement, un nouveau procédé de production d'une protéine cible ayant une modification spécifique d'un site basée sur une liaison carbone-carbone, le procédé comprenant les étapes consistant à : (a) marquer le site à modifier avec une substance ayant un effecteur en ajoutant un acide aminé au site particulier dans la protéine cible; (b) activer le marqueur au niveau du site; et (c) récolter la protéine modifiée en faisant adhérer une fraction de modification post-translationnelle (PTM) sur le marqueur activé. Le procédé de production d'une protéine cible ayant une modification spécifique à un site selon la présente invention non seulement ajoute la modification souhaitée au niveau du site souhaité de la protéine cible, la protéine cible à laquelle une modification a été ajoutée présente également le même effet que la protéine cible présente à l'intérieur de la cellule, et ainsi la présente invention pourrait être utile dans la recherche de la fonction de protéine cible, le criblage de nouveaux médicaments, et analogues. 본 발명은 특정 위치가 변형된(modified) 목적 단백질의 제조방법에 관한 것으로, 보다 구체적으로는 (a) 목적 단백질의 특정 위치에 아미노산을 첨가하여 변형시킬 위치를 반응기를 가지는 물질로 표지하는 단계; (b) 상기 위치의 표지를 활성화시키는 단계; 및 (c) 상기 활성화된 표지에 번역후변형(post translational modification, PTM) 작용기(moiety)를 부착시켜 변형된 단백질을 수득하는 단계를 포함하는 새로운 탄소-탄소 결합 기반 위치 특이적으로 변형된 목적 단백질의 제조방법에 관한 것이다. 본 발명에 따른 위치 특이적으로 변형된 목적 단백질의 제조 방법은 목적 단백질의 원하는 위치에 원하는 변형(modification)을 추가할 뿐만 아니라, 변형이 추가된 목적 단백질이 세포 내에 존재하는 목적 단백질과 동일한 효과를 나타내어 목적 단백질의 기능연구, 신약 스크리닝에 등에 유용하다.