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- 저자 : 고은희 ( Bong Sub Kim (검색결과 2 건)
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김봉섭,권훈정,최명언,고은희,Kim, Bong-Sub,Kwon, Hoon-Jeong,Choi, Myung-Un,Koh, Eun-Hie 생화학분자생물학회 1987 한국생화학회지 Vol.20 No.3
In the previous study of the reaction of the cysteinyl groups of metallothionein (MT) with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) (Korean Biochem. J. (1984) 17, 288-298), it was found that the DTNB titrable cysteinyl groups were markedly affected either by the presence of metal or by the changes of protein conformation. In order to clarify these phenomena, we prepared the a fragment (4-metal cluster) of MT-I and examined the DTNB reaction. The ratio of 1.7 was obtained as the DTNB titrable cysteinyl groups to the bound metal ions. Thus these data support the notion that only one type of cysteinyl groups may interact with DTNB, mainly simply coordinated cysteinyl groups. Furthermore, we investigated the effects of var ous disulfides on the DTNB reaction rates to explore the microenvironment around the cysteinyl groups. Disulfides tested were cystamine, 2-hydroxyethyl disulfide, dimethyl disulfide, and 3,3'-dithiodipropanoic acid. The DTNB reaction rate of MT-I was enhanced by cystamine and 2-hydroxyethyl disulfide, but that of the a fragment was inhibited by both disulfides. Others had no effects on the DTNB reaction of MT. Therefore, the DTNB titrable cysteinyl groups of MT seem to be located in a hydrophilic and negatively charged microenvironment. 메탈로사이오닌과 5,5'-dithiobis(2-nitrobenzoate) (DTNB)의 반응연구에 의하면 (Korean Biochem. J. (1984) 17, 288-289), DTNB에 의하여 적정되는 cysteinyl기의 수는 금속의 존재나 메탈로사이오닌의 구조에 영향을 받는다. 이것을 더 연구하기 위해 메탈로사이오닌의 $\alpha$ fragment를 분리하여 DTNB와 반응시켰다. DTNB로 적정가능한 cysteinyl기의 수와 금속이온수의 비는 1.7이었다. 이것은 메탈로사이오닌 내의 cysteinyl키 중 simply coordinate된 것만이 DTNB와 반응함을 뒷받침 해준다. 더 나아가 cysteinyl 거의 분자내 환경을 알기 위해 disulfide 의 영향을 살펴보았다. Cystamine, 2,2'-dihydroxyethyl disulfide, 3,3'-dithiodipropanoic acid, dimethyl disulfide 동의 disulfide가 사용 되었다. 메탈로사이오닌-I의 경우에는 cystamine과 2,2'-hydroxyethyl disulfide에 의하여 반응속도가 증가하였으나 $\alpha$ fragment의 경우에는 감소하였다. 다른 두 disulfide는 반응속도에 영향을 마치지 않았다. 따라서 DTNB와 반응하는 메탈로사이오닌의 cysteinyl기는 그 미세환경이 음성전하를 띠고 친수성의 위치에 처해 있다고 추정된다.
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5 ' 5 ' - dithiobis ( 2 - nitrobenzoic acid ) 에 의한 메탈로사이오닌 cystein 기의 분자 특징
김봉섭,권훈정,최명언,고은희 ( Bong Sub Kim,Hoon Jeong Kwon,Myung Un Choi,Eun Hie Koh ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.3
In the previous study of the reaction of the cysteinyl groups of metallothionein (MT) with 5,5`-dithiobis(2-nitrobenzoic acid) (DTNB) (Korean Biochem. J. (1984) 17, 288-298), it was found that the DTNB titrable cysteinyl groups were markedly affected either by the presence of metal or by the changes of protein conformation. In order to clarify these phenomena, we prepared the a fragment (4-metal cluster) of MT-I and examined the DTNB reaction. The ratio of 1.7 was obtained as the DTNB titrable cysteinyl groups to the bound metal ions. Thus these data support the notion that only one type of cysteinyl groups may interact with DTNB, mainly simply coordinated cysteinyl groups. Furthermore, we investigated the effects of var ous disulfides on the DTNB reaction rates to explore the micro environment around the cysteinyl groups. Disulfides tested were cystamine, 2-hydroxyethyl disulfide, dimethyl disulfide, and 3,3`-dithiodipropanoic acid. The DTNB reaction rate of MT-I was enhanced by cystamine and 2-hydroxyethyl disulfide, but that of the α fragment was inhibited by both disulfides. Others had no effects on the DTNB reaction of MT. Therefore, the DTNB titrable cysteinyl groups of MT seem to be located in a hydrophilic and negatively charged microenvironment.
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