Gel electrophoresis에 의한 인삼단백질의 구조연구

Abstract

인삼의 여러가지 약효성분들 중 항방사선작용이 있다고 보고된 단백질성분을 분리 정제하여, 그 구조 규명의 첫단계로써 부분정제된 각 분획의 분자량 및 각 분획에 존재하는 subunit의 분자량 측정을 시도하였다. 즉, 표준단백질과 부분정제로부터 얻어진 세개의 인삼분획(GⅠ, GⅡ 및 GⅢ)에 대한 cathodic Disc-Polyacrylamide gel electrophoresis(Disc-PAGE)를 실시하여 표준단백질 및 각 단백질분획 bands의 이동도로부터 분자량을 측정한 후, 따로 Sodium dodecyl sulfate (SDS) 존재하에 Polyacrylamide gel electrophoresis(SDS-PAGE)를 실시하여 표준단백질 및 각 분획에 존재하는 subunit bands의 이동도로부터 subunit 분자량을 측정하였다. Disc-PAGE결과로 나타난 bands의 수효와 분자량은 GⅠ 2개(분자량 : 213,000이상, 55,000), GⅡ 1개(분자량 : 44,000) 및 GⅢ 1개(분자량 : 19,000)로 측정되었으며, SDS-PAGE 결과 각 분획에 존재하는 subunit bands의 수효와 분자량은 GⅠ 4개(subunit 분자량 : 114,000 이상, 27,000, 24,000, 19,000) GⅡ 2개(subunit 분자량 : 46,000, 22,000) 및 GⅢ 1개(subunit 분자량 : 19,000)로 측정되었다. 또한 Silica gel Thin-layer chromatography결과 GⅠ 3개, GⅡ 1개 및 GⅢ 1개의 반점이 검출되었으며, 인삼분획을 SDS로 처리하여 실험한 결과 GⅠ 14개, GⅡ 2개 및 GⅢ 1개의 반점이 검출되었다.;This study was designed to examine the structural property of radiation-protective GⅠnseng proteins. To obtain partially purified ginseng proteins reported to exert anti-radiation effect, Tris-HCl buffer extraction, ammonium sulfate fractionation, CM-cellulose column chromatography, heat inactivation and Sephadex G-75 column chromatography were conducted. After three fractions-GⅠ, CⅡ and GⅢ-were obtained, these three fractions and standard proteins had been analyzed to estimate molecular weights of native proteins by Disc-polyacryl -amide gel electrophoresis and to estimate subunit molecular weights of denatured proteins by SDS-polyacrylamide gel electro-phoresis, respectively. By TLC and Disc-PAGE, GⅠ fraction showed three spots, two bands and molecular weights of two bands were above 213,000 and 55,000 obtained from regression line for standard proteins. GⅡ fraction showed one spot and one band of which molecular weight was 44,000. And GⅢ fraction showed one spot and one band of which molecular weight was 19,000. By SDS-TLC and SDS-PAGE, GⅠ fraction showed four spots and four bands of which subunit molecular weights were above 114,000, 27,000, 24,000 and 19,000 obtained from regression line for standard proteins. GⅡ fraction showed two spots and two bands of which subunit molecular weights were 46,000 and 22,000. And GⅢ fraction showed one spot and one band of which subunit molecular weight was 19,000.