Protein crystallization in hydrogel beads.
Protein crystallization in hydrogel beads.
复制标题
水凝胶珠中的蛋白质结晶。
DOI：
10.1107/s0907444905021566
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
M. Sleutel
中科院分区：
文献类型：
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作者：
R. Willaert;I. Zegers;L. Wyns;M. Sleutel
The use of hydrogel beads for the crystallization of proteins is explored in this contribution. The dynamic behaviour of the internal precipitant, protein concentration and relative supersaturation in a gel bead upon submerging the bead in a precipitant solution is characterized theoretically using a transient diffusion model. Agarose and calcium alginate beads have been used for the crystallization of a low-molecular-weight (14.4 kDa, hen egg-white lysozyme) and a high-molecular-weight (636.0 kDa, alcohol oxidase) protein. Entrapment of the protein in the agarose-gel matrix was accomplished using two methods. In the first method, a protein solution is mixed with the agarose sol solution. Gel beads are produced by immersing drops of the protein-agarose sol mixture in a cold paraffin solution. In the second method (which was used to produce calcium alginate and agarose beads), empty gel beads are first produced and subsequently filled with protein by diffusion from a bulk solution into the bead. This latter method has the advantage that a supplementary purification step is introduced (for protein aggregates and large impurities) owing to the diffusion process in the gel matrix. Increasing the precipitant, gel concentration and protein loading resulted in a larger number of crystals of smaller size. Consequently, agarose as well as alginate gels act as nucleation promoters. The supersaturation in a gel bead can be dynamically controlled by changing the precipitant and/or the protein concentration in the bulk solution. Manipulation of the supersaturation allowed the nucleation rate to be varied and led to the production of large crystals which were homogeneously distributed in the gel bead.
Miyoshi, K.：“rRNA 成熟/核糖体亚基组装所需的反式作用因子基因的转录与核糖体蛋白基因协调调节，并涉及酿酒酵母中的 Rap1”，《核酸研究》31・7。
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