CONTROL OF NUCLEATION OF PROTEIN CRYSTALS

CONTROL OF NUCLEATION OF PROTEIN CRYSTALS
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DOI:
10.1002/pro.5560031003
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发表时间:
1994-10-01
期刊:
影响因子:
8
通讯作者:
SARIDAKIS, E
SARIDAKIS, E
中科院分区:
生物学3区
文献类型:
--
作者:
BLOW, DM;CHAYEN, NE;SARIDAKIS, E

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当标准技术得到许多小晶体或孪生晶体时,可能需要控制成核以获得大蛋白质晶体的可靠供应。非均相成核可以通过使用精细过滤器来控制,并通过在石蜡油下工作来消除空气中的污染物。与支撑容器的接触面积也有重要的影响。溶菌酶的异质成核剂(前面已经确定)已被证明对羧肽酶G有效(2)控制均质成核(以前通过在不同时间的孵育后稀释成核样品来证明)也可以通过在1温度下孵育样品来实现,在1温度下可以发生成核,然后改变温度到有生长但没有成核的条件。
Control of nucleation may be needed to obtain a reliable supply of large protein crystals, when standard techniques give many small or twinned crystals. Heterogeneous nucleation may be controlled by the use of fine filters, with the elimination of airborne contaminants by working under paraffin oil. The area of contact with the supporting vessel also has an important effect. A heterogeneous nucleant for lysozyme (identified earlier) has been shown to be effective for carboxypeptidase G(2) Control of homogeneous nucleation (previously demonstrated by dilutions of a nucleating sample after various times of incubation) may also be achieved by incubating a sample at 1 temperature, where nucleation can occur, and changing the temperature to conditions where there is growth but no nucleation.