公开(公告)号：KR1020090121441A

公开(公告)日：2009-11-26

申请号：KR1020080047349

申请日：2008-05-22

Applicant: 국민대학교산학협력단

Inventor： 유연규 ,  이원규 ,  이상엽

IPC: G01N33/68 ,  G01N33/53 ,  G01N33/48

Abstract: PURPOSE: A method for measuring interaction between CK2α and Nopp140 and a method for suppressing CK2 activation are provided to massively detect a compound without expensive cell culture stage. CONSTITUTION: A method for measuring interaction of CK2α and Nopp140 comprises: a step of fixing GST-CK2α protein on a multi-well plate; a step of treting Nopp140 protein; and a step of measuring the amount of Nopp140 bound to GST-CK2α. A method for screening an interaction regulating material of CK2 and Nopp140 comprises: a step of reacting CK2 subunit and a candidate of interaction regulation of CK2 and Nopp140 and InsP6(inositolhexakisphosphate); and a step of treating Nopp140 to a reacted material.

Abstract translation: 目的：提供一种测定CK2α与Nopp140之间相互作用的方法及抑制CK2活化的方法，以大量检测化合物，而无需昂贵的细胞培养阶段。 构成：测定CK2α和Nopp140相互作用的方法包括：将多肽板上固定GST-CK2α蛋白的步骤; 处理Nopp140蛋白的一个步骤; 以及测定与GST-CK2α结合的Nopp140的量的步骤。 筛选CK2和Nopp140的相互作用调节材料的方法包括：使CK2亚基与CK2和Nopp140和InsP6（肌醇六磷酸酯）的相互作用调节候选物反应的步骤; 以及将Nopp140处理成反应材料的步骤。

公开(公告)号：KR1020080071330A

公开(公告)日：2008-08-04

申请号：KR1020070009411

申请日：2007-01-30

Applicant: 국민대학교산학협력단

Inventor： 유연규 ,  이원규

IPC: G01N33/68 ,  G01N33/53 ,  G01N33/48

Abstract: A screening method of a regulating agent for the signal transduction in BLT2 as a receptor of leukotriene B4 is provided to reduce the screening costs by eliminating a cell culture step by using purified protein and screening devices, so that candidate new drug substances for treating inflammatory disease are screened rapidly and inexpensively. A screening method of a regulating agent for the signal transduction in BLT2 comprises the steps of: (1) preparing BLT2 protein or protein containing an intracellular loop 3(iL3) region of BLT2; (2) preparing alpha subunit protein of G protein(Galpha); (3) reacting the prepared protein of step (1) with the protein of step (2) in the presence of a testing compound, and measuring the interaction between proteins; and (4) comparing the measured interaction with that of a contrast which is measured in the absence of the testing compound, wherein the BLT2 protein is a fusion protein of protein purification-facilitating regions including N-terminal T7 tag and C-terminal His-tag and a BLT2 region; and the protein containing an intracellular loop 3(iL3) region of BLT2 is a fusion protein of a protein purification-facilitating region such as GST(glutathione-S-transferase) and a BLT2 iL3 region. Further, the alpha subunit protein of G protein(Galpha) is alpha subunit protein of Gi3 protein.

Abstract translation: 提供BLT2作为白细胞三烯B4受体的信号转导调节剂的筛选方法，通过使用纯化的蛋白质和筛选装置消除细胞培养步骤来降低筛选成本，从而用于治疗炎性疾病的候选新药物 被快速和廉价地筛选。 BLT2信号转导调节剂的筛选方法包括以下步骤：（1）制备含有BLT2细胞内环3（iL3）区域的BLT2蛋白或蛋白质; （2）制备G蛋白（Galpha）的α亚基蛋白; （3）在试验化合物的存在下使步骤（1）的制备的蛋白质与步骤（2）的蛋白质反应，并测量蛋白质之间的相互作用; （4）将测定的相互作用与不存在测试化合物时所测量的相互作用进行比较，其中BLT2蛋白是蛋白质纯化促进区的融合蛋白，其包括N末端T7标记和C-末端His- 标签和BLT2区域; 并且含有BLT2的细胞内环3（iL3）区域的蛋白质是蛋白质纯化促进区域如GST（谷胱甘肽-S-转移酶）和BLT2 iL3区域的融合蛋白。 此外，G蛋白（Galpha）的α亚基蛋白是Gi3蛋白的α亚基蛋白。