CALCULATION OF PROTEIN EXTINCTION COEFFICIENTS FROM AMINO-ACID SEQUENCE DATA
CALCULATION OF PROTEIN EXTINCTION COEFFICIENTS FROM AMINO-ACID SEQUENCE DATA
复制标题
DOI：
10.1016/0003-2697(89)90602-7
复制
发表时间：
1989-11-01
影响因子：
2.9
通讯作者：
VONHIPPEL, PH
中科院分区：
文献类型：
--
作者：
GILL, SC;VONHIPPEL, PH
溶液中蛋白质-蛋白质和蛋白质-配体相互作用的定量研究需要准确测定蛋白质浓度。通常，对于仅以“分子生物学”量可用的蛋白质，很难或不可能对蛋白质的摩尔消光系数进行准确的实验测量。然而，没有一个可靠的值，这个参数，不能确定蛋白质浓度的通常紫外光谱手段。幸运的是，氨基酸残基序列和原聚体分子量（因此也是氨基酸组成）的知识通常可以通过DNA序列获得，对于大多数这样的蛋白质，DNA序列通常是准确已知的。本文提出了一种计算精确（to . ±. 5%，在大多数情况下）的摩尔消光系数的蛋白质在280 nm，简单地从知识的氨基酸组成。该方法是校准对18个“正常”的球状蛋白质的摩尔消光系数是准确的，和假设的方法，以及它的局限性，进行了讨论。
Quantitative study of protein-protein and protein-ligand interactions in solution requires accurate determination of protein concentration. Often, for proteins available only in "molecular biological" amounts, it is difficult or impossible to make an accurate experimental measurement of the molar extinction coefficient of the protein. Yet without a reliable value of this parameter, one cannot determine protein concentrations by the usual uv spectroscopic means. Fortunately, knowledge of amino acids residue sequence and protomer molecular weight (and thus also of amino acid composition) is generally available through the DNA sequence, which is usually accurately known for most such proteins. In this paper we present a method for calculating accurate (to .+-. 5% in most cases) molar extinction coefficients for proteins at 280 nm, simply from knowledge of the amino acid composition. The method is calibrated against 18 "normal" globular proteins whose molar extinction coefficients are accurately known, and the assumptions underlying the method, as well as its limitations, are discussed.