ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY OF MOLECULAR VARIANTS OF A [2FE-2S] FERREDOXIN
ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY OF MOLECULAR VARIANTS OF A [2FE-2S] FERREDOXIN
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DOI：
10.1006/bbrc.1995.1714
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发表时间：
1995-05-25
影响因子：
3.1
通讯作者：
MEYER, J
中科院分区：
文献类型：
--
作者：
PETILLOT, Y;GOLINELLI, MP;MEYER, J
巴氏梭菌[2Fe-2S]铁氧还蛋白是一种同源二聚体蛋白，每个亚基含有一个[2Fe-2S]簇。在先前的研究中，位置11、14、24、56和60的五个半胱氨酸残基已突变为丝氨酸或丙氨酸。野生型铁氧还蛋白及其几种分子变体现在已经通过电喷雾电离质谱法进行了分析。在负离子检测模式下，根据所使用的输注溶剂，在所有情况下均检测到归因于脱辅基蛋白、单体全蛋白和二聚体全蛋白的分子峰。数据证实了预期突变的存在，表明所有这些蛋白质每个亚基含有一个[2Fe-2S]簇，并表明这些铁氧还蛋白的二聚体结构可以在电喷雾电离条件下保留。本研究建立了电喷雾离子化质谱分析含有不稳定金属簇的寡聚蛋白质的功率。(C)出版社：Academic Press
The [2Fe-2S] ferredoxin from Clostridium pasteurianum is a homodimeric protein of which each subunit contains one [2Fe-2S] cluster. In previous investigations, the five cysteine residues in positions 11, 14, 24, 56 and 60 had been mutated into serine or alanine. The wild type ferredoxin and several of its molecular variants have now been analyzed by electrospray-ionization mass spectrometry. In the negative-ion detection mode, depending on the infusion solvent used, molecular peaks attributable to the apoprotein, to the monomeric holoprotein, and to the dimeric holoprotein were detected in all cases. The data confirmed the presence of the expected mutations, showed that all of these proteins contain one [2Fe-2S] cluster per subunit, and indicated that the dimeric structure of these ferredoxins could be retained in the conditions of the electrospray ionization. This investigation establishes the power of electrospray-ionization mass spectrometry for the analysis of oligomeric proteins containing labile metal clusters. (C) 1995 Academic Press, Inc.