一些与固定化核苷酸亲和色谱相关的参数。
Some parameters relevant to affinity chromatography on immobilized nucleotides.
作者信息
Lowe C R, Harvey M J, Craven D B, Dean P D
出版信息
Biochem J. 1973 Jul;133(3):499-506. doi: 10.1042/bj1330499.
Abstract
- The suitability of cellulose and Sepharose as supports for affinity chromatography of two groups of cofactor-linked enzymes, dehydrogenases and kinases, was examined. Sepharose was found to be superior. 2. The selective capacities of the columns were measured by frontal analysis and are discussed in relation to the nucleotide contents. 3. The effect of various concentrations of enzyme and of non-specific protein on the performance of the affinity columns, and the effects of equilibration time, flow rate, sample volume and dilution of the nucleotide were examined. 4. The effect of interposing polymethylene and polyglycine extension arms between the matrix backbone and the nucleotide was investigated for several cofactor-dependent enzymes. Maximum binding was observed with an extension arm 0.8-1nm long.
摘要
- 研究了纤维素和琼脂糖作为两类与辅因子相连的酶（脱氢酶和激酶）亲和色谱支持物的适用性。结果发现琼脂糖更具优势。2. 通过前沿分析法测定了柱的选择性容量，并结合核苷酸含量进行了讨论。3. 考察了不同浓度的酶和非特异性蛋白质对亲和柱性能的影响，以及平衡时间、流速、样品体积和核苷酸稀释的影响。4. 针对几种依赖辅因子的酶，研究了在基质主链和核苷酸之间插入亚甲基和聚甘氨酸延伸臂的效果。观察到延伸臂长度为0.8 - 1纳米时结合力最大。
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