蛋白质顺序分析

又称 蛋白质的一级结构分析.对组成蛋白质多 肽链的氨基酸排列顺序进行测定的方法.此法因１９５４年英国化学家Ｆ. 桑格成功地测定了 牛胰岛素的氨基酸排列顺序而奠定了基础.此法大致包括以下步骤.一、根据蛋白质的氨基（Ｎ）末端残基或 羧基（Ｃ）末端残基的 摩尔数和蛋白质的分子量，确定蛋白质的 多肽链数.如果蛋白质由几条不同的多肽链组成，就须设法先把这些多肽链拆分开来，再测定各条多肽链的氨基酸顺序.二、将分离、提纯多肽链的部分样品完全水解，用离子交换 层析法测定肽链的氨基酸组成，并算出各种氨基酸的分子比.三、应用有关的 化学试剂确定多肽链Ｎ末端和Ｃ末端的 氨基酸残基.四、用几种不同的方法将肽链样品断裂成几套肽段，并用层析法将这些肽段分开.五、用适当的降解法确定各肽段的氨基酸顺序.六、根据几套肽段氨基酸顺序之间的重叠情况，就可推断出整条多肽链的氨基酸顺序.上述方法工作量大，费时费事，后来使用 氨基酸自动分析仪，效率大为提高.目前采用从ＤＮＡ的核苷酸顺序推测多肽链的氨基酸顺序的方法进行测定，比较快速、简便，已被广泛应用. Also known as a structural analysis of proteins. For the composition of the protein polypeptide chain of amino acids sequences were determined approach. This method because in 1954 the British chemist F. Sanger successfully determined the amino acid sequence of bovine insulin and laid the basis. This method generally includes the following steps. 1, according to the protein's amino (N)-terminal residue or the carboxyl (C)-terminal residues of the mole numbers and protein molecular weight, determine the number of protein polypeptide chain. If the protein has several composed of different polypeptide chains, it should first of these to be trying to split off from the polypeptide chain, and then measured the polypeptide chain of each amino acid sequence. Second, the separation, purification polypeptide chain part of the sample completely hydrolyzed, using ion-exchange chromatography determination of amino acid composition of peptide chain, and calculate the molecular ratio of various amino acids. 3, application of chemical reagents to determine the polypeptide chain N-terminal and C-terminal amino acid residues. 4, with several different ways polypeptide chain broken into several sets of peptide samples and use chromatography to separate these peptides. 5, using the appropriate method to determine the degradation of the amino acid sequence of each peptide. 6, according to several sets of peptide amino acid sequence of the overlap between the , we can infer that the entire amino acid sequence of the polypeptide chain. the above method workload, time and effort, then use the amino acid automatic analyzer, efficiency greatly improved. currently used to speculate from the DNA nucleotide sequence of the polypeptide chain of amino acids sequence determination methods, comparing fast, simple and has been widely used.