생명과학회지 (Journal of Life Science)

  • 제10권3호
  • /
  • Pages.279-285
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  • 2000
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지

온도변화에 의한 cAMP 수용성 단백질(CRP)의 구조

Study on the structure of cAMP receptor protein(CRP) by temperature change

  • 발행 : 2000.06.01
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초록

cAMP 수용성 단백질인 CRP는 Escherichia coli에서 대사와 관련된 유전자의 전사를 조절한다. 본 연구는 야생형과 돌연변이 CRP 단백질의 열적 안정성과 온도에 따른 단백질의 구조변화를 관찰하기 위 하여 proteolytic digestion, UV spectrophotometer, CD spectrapolarimeter 등의 방법을 사용하였다. cAMP가 없을 때에는 야생형, S83G, S128A CRP가 열적 안정성에서 큰 차이를 보이지 않았지만, cAMP가 존재할 때 야생형 CRP가 다른 돌연변이 CRP보다 열적으로 더욱 안정함을 보였다. 그리고 protease digestion 실험을 통하여 높은 온도에서 cAMP의 존재와 무관하게 돌연변이 CRP에서 단백질 의 변성으로 인한 절단된 단백질띠를 관찰할 수 있었다. 그리고 55$^{\circ}C$에서 측정한 CD 스펙트럼에서 단백 질의 2차 구조인 $\alpha$-helix 구조가 부분적으로 파괴되었음이 관찰되었다.

CRP (cyclic AMP receptor protein) regulate transcription of catabolite-sensitive genes in Escherichia coli. Wild-type and mutant CRP (S83G and S128A) proteins were used to measure the thermal stability and the temperature-dependent structural change by proteolytic digestion, UV spectrophotometer and CD spectrapolarimeter. The result indicated that wild-type CRP was more thermally stable than the mutant CRPs in the presence of cAMP. At a low temperature, wild-type CRP with cAMP was more sensitive to subtilisin than the mutant CRPs. At a high temperature, there was no difference of sensitivity to subtilisin among wild-type, S83G and S128A CRPs. CD spectra suggested that the secondary structure of CRP was destroyed partially at a high temperature.

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참고문헌

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