Korean Journal of Crystallography (한국결정학회지)

Korean Crystallographic Association (한국결정학회)
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Evaporation Rate in Protein Crystallization Via Vapor Diffusion can be Controlled through a Simple Multistep-concentration Setting in Capillaries

  • Lee, Min-Nyung (School of Biological Sciences, Chungbuk National University) ;
  • Chung, Yong-Je (School of Biological Sciences, Chungbuk National University)
  • Published : 2003.06.01
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Abstract

A simple multistep-concentration setting in capillaries was used to control the water-evaporation rate in vapor-diffusion protein crystallization. In the method used, a variety of evaporation rate curves were obtained by using the secondary precipitant solution referred to as “regulatory solution”, which is not directly exposed to the protein solution. The curves were applied to the crystallization of lysozyme as a model protein. The results clearly showed that crystal growth is dependent on the evaporation rate. Especially, the decoupling curves in which precipitant concentration in protein solutions increases to a certain point and then decreases to the equilibrium concentration gave the best crystals.

증기-확산 단백질 결정화의 물-증발 속도를 조절하기 위해서, 간결한 모세관 내의 다중-단계 농도 장착법 (multistep-concentration setting in capillaries) 이 이용되었다. “조절 용액 (regulatory solution)”이라고 일컬어지는 2차 침전 용액이 이용되어, 이 방법으로 다양한 증발 속도 곡선들이 얻어졌다. 이때, 조절 용액은 단백질 용액에 직접적으로 노출되지 않는다. 모델 단백질인 lysozyme의 결정화에 이 그래프들이 적용되었다. 결정 성장은 증발속도에 달려있다는 것을 실험 결과들이 명백하게 보여주었다. 특히나. 단백질 용액의 침전 농도가 어떤 점까지 증가하다가 평형 농도로 줄어드는 decoupling 곡선이 가장 좋은 결정들을 만들어냈다.

Keywords

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