The primary structure of thioredoxin from Chromatium vinosum determined by high-performance tandem mass spectrometry.
The primary structure of thioredoxin from Chromatium vinosum determined by high-performance tandem mass spectrometry.
复制标题
通过高性能串联质谱法测定 Chromatium vinosum 中硫氧还蛋白的一级结构。
DOI：
10.1021/bi00379a001
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Biemann,K
中科院分区：
文献类型：
--
作者：
Johnson,RS;Biemann,K
麻省理工学院化学系，剑桥，马萨诸塞州02139摘要：用高效串联质谱法测定了从vinosum中分离的氧化还原蛋白硫氧还蛋白的一级结构，并对胰蛋白酶消化产生的14个肽（长度从2到18个氨基酸不等）和金黄色葡萄球菌蛋白酶产生的几个肽进行了测序。用质谱法测定后一消化产物中肽的分子量，并根据与大肠杆菌硫氧还蛋白相当的同源性，对色氨酸肽进行了正确的排列。最后，用质谱法测定了硫氧还蛋白的分子量为11 748.0，与该序列计算的11 750.2符合得很好。虽然用质谱法很难确定，但可以鉴定出5种亮氨酸和3种异亮氨酸，剩下8种未区分。
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 Received November 26, 1986; Revised Manuscript Received January 7, 1987 abstract: The primary structure of thioredoxin, a redox protein isolated from Chromatium vinosum, was determined by high-performance tandem mass spectrometry, which permitted sequencing of the 14 peptides (ranging in length from 2 to 18 amino acids) generated by digestion with trypsin and of several peptides produced by Staphylococcus aureus protease. The mass spectrometrically determined molecular weights of the peptides from the latter digest were used to properly align the tryptic peptides, which could alsobe accomplished on the basis of the considerable homology with Escherichia coli thioredoxin. Finally, the molecular weight of the Chromatium thioredoxin was determined by mass spectrometry and foundto be 11 748.0, in good agreement with 11 750.2 calculated for the proposed sequence. Although it was difficult to establish by mass spectrometry, five leucines and three isoleucines could be identified, leaving only eight undifferentiated.