Methods for identifying proteins by using partial sequences.
Methods for identifying proteins by using partial sequences.
复制标题
使用部分序列鉴定蛋白质的方法。
DOI：
--
复制
发表时间：
1979
影响因子：
11.1
通讯作者：
B. C. Orcutt
中科院分区：
文献类型：
--
作者：
M. O. Dayhoff;B. C. Orcutt
描述了利用部分序列分析信息鉴定蛋白质片段的方法。如果蛋白质序列已知，通常只知道7个氨基酸残基（不一定是连续的）的身份和位置，通过与所有已知序列的数据文件进行比较，就可以确定一个片段。部分序列是通过极其敏感的微测序程序获得的。组织用氨基酸孵育，其中一种或多种氨基酸具有明显的放射性标记。分离出感兴趣的蛋白质，并进行测序实验，以确定大约30个残基的nh2末端段的放射性位置。我们推导并研究了一个方程，用于找到与任何放射性模式唯一匹配的概率。据此，我们提出一个新的策略。在一次孵育中，用每种同位素标记几种氨基酸。大多数信息包含在由不同标签（包括无标签）区分的残基占据大约相等数量位置的模式中。片段的氨基酸组成通常不会事先知道。标记残基预计占据36%的位置，足以有98%的机会成功地独特地表征任何人类片段。这种策略将允许从单个组织孵育中鉴定大多数蛋白质。数学上的讨论是一般的，适用于序列的任何段，也适用于用任何方法得到的序列。改进的鉴定程序应加快蛋白质表达和功能信息的积累。
Methods for the identification of a protein segment by using the information from partial sequence analyses are described. If the protein sequence is known, a segment can usually be identified with confidence through comparison with the data file of all known sequences when the identity and position of only seven amino acid residues (not necessarily contiguous) are known. Partial sequences are obtained from extremely sensitive microsequencing procedures. Tissue is incubated with amino acids, one or more of which are distinctively radiolabeled. Proteins of interest are isolated and a sequenator experiment performed to locate the positions of radioactivity in an NH(2)-terminal segment of approximately 30 residues. We derive and investigate an equation for the probability of finding a unique match to any pattern of radioactivity. From this we suggest a new strategy. In one incubation, several amino acids are labeled with each kind of isotope. The most information is contained in patterns in which approximately equal numbers of positions are occupied by residues distinguished by different labels (including no label). The amino acid composition of the segment will typically not be known in advance. Labeling residues expected to occupy 36% of the positions suffices for a 98% chance of success in uniquely characterizing any human segment. Such a strategy will permit the identification of most proteins from a single tissue incubation. The mathematical discussion is general and applies to any segment from a sequence and to sequences obtained by any method. Improved identification procedures should expedite the accumulation of information on the expression and function of proteins.