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One‐step purification of the NADH dehydrogenase fragment of the Escherichia coli complex I by means of Strep ‐tag affinity chromatography
Author(s) -
Bungert Stefanie,
Krafft Bianca,
Schlesinger Ramona,
Friedrich Thorsten
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01341-1
Subject(s) - escherichia coli , flavin mononucleotide , affinity chromatography , nadh dehydrogenase , oxidoreductase , biochemistry , chemistry , fragment (logic) , dehydrogenase , stereochemistry , flavin group , enzyme , protein subunit , computer science , programming language , gene
The proton‐pumping NADH:ubiquinone oxidoreductase, also called complex I, is the first energy‐transducing complex of many respiratory chains. Complex I of Escherichia coli can be split into three fragments. One of these fragments, the soluble NADH dehydrogenase fragment, represents the electron input part of complex I. It comprises the subunits NuoE, F and G and harbors one flavin mononucleotide and up to six iron‐sulfur clusters. Here, we report the one‐step purification of this fragment by means of affinity chromatography on StrepTactin. This was achieved by fusing the Strep ‐tag II peptide to the C‐terminus of NuoF or NuoG. Fusion of this peptide to the N‐terminus of either NuoE or NuoF disturbed the assembly of the NADH dehydrogenase fragment.