Yamamoto A, Toda H, Sakiyama F
Institute for Protein Research, Osaka University.
J Biochem. 1989 Oct;106(4):552-4. doi: 10.1093/oxfordjournals.jbchem.a122892.
The "vapor-phase" hydrazinolysis method was devised for the microdetermination of the carboxyl-terminal residue of a protein. With this method, a polypeptide sample is degraded with vaporized hydrazine. The optimum conditions for hen egg-white lysozyme were established to be 2 to 4 h at 90 or 100 degrees C, the recovery of the carboxyl-terminal leucine being about 70%. With this vapor-phase method, side reactions are reduced and the time of hydrazinolysis is shortened. The limit of quantitation for the carboxyl-terminus of a protein is about 50 pmol, as judged so far with hen egg-white lysozyme. The carboxyl-termini of several proteins were determined using this novel procedure.
“气相”肼解方法是为微量测定蛋白质的羧基末端残基而设计的。用这种方法，多肽样品用汽化肼降解。已确定鸡蛋清溶菌酶的最佳条件是在90或100℃下反应2至4小时，羧基末端亮氨酸的回收率约为70%。用这种气相方法，副反应减少，肼解时间缩短。就目前用鸡蛋清溶菌酶判断的情况而言，蛋白质羧基末端的定量限约为50皮摩尔。使用这种新方法测定了几种蛋白质的羧基末端。
