단백질 독소 리신의 생화학적 분석법 정립에 관한 연구
A Study on the Establishment of Biochemical Analysis of Ricin Protein toxin
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초록
리신은 피마자(Ricinus communis, 아주까리)씨 전체 질량의 1∼5% 함유되어 있는 단백질 독소로, 세포 내 단백질 합성을 억제함으로써 독성을 나타낸다. 보툴리눔 독소에 비해 독성은 1,000배 정도 약하지만, 재료를 쉽게 구할 수 있고 대량생산이 용이하다는 점에서 테러 사용 가능성이 높은 물질이다. 체내 흡수 시 증상 발현이 빠르고 치명적이므로 의심 환경시료 내 리신 존재 유무의 신속하고 정확한 분석은 생물 상황 대응을 위해 필수적인 과정이다. 본 연구에서는 리신의 생화학적 특성 분석 및 검출을 위하여 Western blot 분석법을 구축하였으며, 이를 통해 리신 단백질의 분자량 확인 및 정성분석이 가능함을 확인하였다. 특히, 단백질 전기영동을 통해 환원(reduced) 및 비환원(non-reduded) 조건에서의 리신 내 이황화 결합(disulfide bond)으로 연결된 2개의 소단위체(subunit)를 확인하였으며, 각 subunit의 항체(Ricin A-chain, B-chain antibody)를 이용하여 subunit 단위에서의 정성분석이 가능함을 확인하였다. 본 연구를 통한 리신의 정밀분석법 구축으로 군 내 생물 대응 능력 향상에 기여할 수 있을 것으로 기대한다.
Abstract
Ricin is a protein toxin contained 1-5% in Ricinus communis(Castor bean) seeds, which causes toxicity by inhibiting protein synthesis in human cells. Although the toxicity is 1,000 times weaker than that of botulinum toxins, it is a highly potential materials to be used for terrorism because raw material are easily available and mass production of toxins is easy. When Ricin is absorbed into the body, symptoms are fast and fatal compared to biological agents. So quick and accurate analysis of the presence or absence of ricin in a suspected biological situation is an essential process for response and follow-up. In this study, to identify the biochemical properties of ricin, we established western blot analysis method which can confirmed the molecular weight and qualitative analysis of ricin protein. In particular, two subunits connected by disulfide bonds in ricin under reduced and non-reduced conditions were identified by protein electrophoresis method. And we checked that the qualitative analysis in subunit level of ricin protein was possible using antibodies(Ricin A-chain, B-chain antibody) of each subunit. It is expected that the establishment of analysis method of ricin through this study will contribute to the improvement of biological response capabilities.
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