Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Synthesis and Secretion of Mutant Mannose-Binding Lectin

돌연변이 Mannose-binding Lectin 합성과 세포 병리적 연구

  • Jang, Ho-Jung (Department of Clinical Laboratory Science, Dong-Eui University) ;
  • Chung, Kyung Tae (Department of Clinical Laboratory Science, Dong-Eui University)
  • 장호정 (동의대학교 임상병리학과) ;
  • 정경태 (동의대학교 임상병리학과)
  • Received : 2013.02.19
  • Accepted : 2013.03.26
  • Published : 2013.03.30
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Abstract

Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.

선천성 면역은 감염성 매개체를 자기(self)로부터 변별할 수 있다. 선천성 면역은 감염 초기에 숙주인 자기를 보호하는 인식분자와 효과인자들로 구성되어 있다. Mannose 결합 렉틴(Mannose-binding lectin, MBL)은 $Ca^{2+}$ 의존형 렉틴에 속하며, 콜라겐 유사 domain을 함유하는 C-type 렉틴으로 선천성 면역의 중요한 분자이다. 혈액내 낮은 MBL 농도는 면역결핍 증후군을 나타내며 감염에 대한 심각한 위험성을 초래한다. 사람의 MBL 결핍증은 coding 영역의 돌연변이에 의해 나타나며, 이 돌연변이의 영향을 연구하기 위해 쥐의 상동성 유전자인 MBL-A를 이용하고 있다. 돌연변이 MBL의 기능적, 세포 생리적 연구를 위해 선행연구에서 rat wild type MBL-A 유전자를 클로닝하였으며, 본 연구에서 이 유전자에 콜라겐 유사 domain에서 발견된 세 가지 돌연변이, R40C, G42D, G45E를 site-directed mutagenesis 방법으로 모두 도입하였다. 세 가지 돌연변이가 존재하는 MBL 단백질은 정상 MBL과 마찬가지로 세포 내에서 정상적으로 발현되었으며, 여전히 렉틴 기능을 가지고 있었다. 이는 세 가지 돌연변이가 렉틴 기능을 나타내는 C-말단 쪽의 carbohydrate recognition domain에는 구조적으로, 또한 기능적으로도 영향을 미치지 않는다는 결과이다. 그러나 이 돌연변이는 MBL 단백질이 세포 밖으로 분비되는 것을 방해하였으며, 그 결과로 소포체 내에 잔류하여 소포체 망상구조(endoplasmic reticulumn network)에 커다란 손상을 주며 비정상적인 형체를 초래하였다. 이 같은 결과는 돌연변이 MBL에 의해 나타난 세포 내 병리현상의 새로운 발견으로 향후 MBL의 구조 형성과 분비 연구에 기여를 할 것으로 생각된다.

Keywords

References

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