公开(公告)号：KR100451594B1

公开(公告)日：2004-10-08

申请号：KR1020020006554

申请日：2002-02-05

Applicant: 한국과학기술연구원

Inventor： 유영숙 ,  남희선 ,  반은미 ,  유은아 ,  이세윤

IPC: C12Q1/48

Abstract: PURPOSE: A determination method of protein phosphorylation reaction by enzyme activity is provided, thereby easily and rapidly analyzing the protein phosphorylation reaction. CONSTITUTION: A determination method of protein phosphorylation reaction by enzyme activity comprises the steps of: determining the optimal pH and concentration of a buffer solution for phosphorylation reaction of myelin basic protein(MBP) by extracellular signal-regulated kinase(ERK); adding the optimal concentration of compound to the buffer solution in order to inhibit the adsorption of peptide to the inner wall of a capillary tube and increase the detection sensitivity; determining MBP peptide from the active region of MBP peptide by ERK, -PRTP- as a center for effectively carrying out the phosphorylation reaction by ERK under the same condition; detecting the phosphorylation reaction of MBP peptide by ERK using a capillary tube electrophoresis using the buffer solution; and additionally confirming the phosphorylation reaction detected using a matrix-supported laser removing/ionizing mass analyzer.

Abstract translation: 目的：提供一种通过酶活性进行蛋白质磷酸化反应的测定方法，从而可以容易且快速地分析蛋白质磷酸化反应。 构成：通过酶活性确定蛋白质磷酸化反应的方法包括以下步骤：通过细胞外信号调节激酶（ERK）确定用于磷酸化髓磷脂碱性蛋白（MBP）反应的缓冲溶液的最佳pH和浓度; 将最佳浓度的化合物添加到缓冲液中以抑制肽吸附到毛细管内壁并提高检测灵敏度; 通过ERK以-PRTP-从MBP肽的活性区域测定MBP肽作为用于在相同条件下通过ERK有效进行磷酸化反应的中心; 使用缓冲溶液使用毛细管电泳检测由ERK检测MBP肽的磷酸化反应; 并且另外确认使用基质支持的激光去除/电离质量分析仪检测到的磷酸化反应。

公开(公告)号：KR1020030066217A

公开(公告)日：2003-08-09

申请号：KR1020020006554

申请日：2002-02-05

Applicant: 한국과학기술연구원

Inventor： 유영숙 ,  남희선 ,  반은미 ,  유은아 ,  이세윤

IPC: C12Q1/48

Abstract: PURPOSE: A determination method of protein phosphorylation reaction by enzyme activity is provided, thereby easily and rapidly analyzing the protein phosphorylation reaction. CONSTITUTION: A determination method of protein phosphorylation reaction by enzyme activity comprises the steps of: determining the optimal pH and concentration of a buffer solution for phosphorylation reaction of myelin basic protein(MBP) by extracellular signal-regulated kinase(ERK); adding the optimal concentration of compound to the buffer solution in order to inhibit the adsorption of peptide to the inner wall of a capillary tube and increase the detection sensitivity; determining MBP peptide from the active region of MBP peptide by ERK, -PRTP- as a center for effectively carrying out the phosphorylation reaction by ERK under the same condition; detecting the phosphorylation reaction of MBP peptide by ERK using a capillary tube electrophoresis using the buffer solution; and additionally confirming the phosphorylation reaction detected using a matrix-supported laser removing/ionizing mass analyzer.

Abstract translation: 目的：提供通过酶活性测定蛋白质磷酸化反应的方法，从而容易且快速地分析蛋白质磷酸化反应。 构成：通过酶活性测定蛋白质磷酸化反应的方法包括以下步骤：通过细胞外信号调节激酶（ERK）测定髓磷脂碱性蛋白（MBP）的磷酸化反应缓冲溶液的最佳pH和浓度; 向缓冲溶液中加入最佳浓度的化合物，以抑制肽对毛细管内壁的吸附，提高检测灵敏度; 通过ERK，-PRTP-作为中心，通过ERK在相同条件下有效地进行磷酸化反应，从MBP肽的活性区确定MBP肽; 通过使用缓冲溶液的毛细管电泳检测ERK的MBP肽的磷酸化反应; 并且另外确认使用基质支持的激光去除/电离质量分析仪检测的磷酸化反应。