Application of cell-free translation systems to studies of cofactor binding proteins.
To develop applications of in vitro cell-free translation systems for production and characterization of cofactor binding proteins, we investigate the production of apo- or holo-forms of Flavin Mono Nucleotide (FMN)-binding protein from Desulfovibrio vulgaris (Miyazaki F) and purified them. The redox potential analysis and measurements of UV-, visible, and fluorescent spectra of reconstructed holo-protein showed that the FMN correctly bound to the FMN binding site. On the other hand, contrary to our expectation, we found that the apo-protein formed a dimer structure and the incorporation of the FMN led the conformational alterations of the protein. These studies demonstrate the utility of cell-free translation systems to analyses of cofactor-binding proteins.
Abe
Masato
M
Department of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama 790-8577, Japan.
Hori
Hiroyuki
H
Nakanishi
Takeshi
T
Arisaka
Fumio
F
Ogasawara
Tomio
T
Sawasaki
Tatsuya
T
Kitamura
Masaya
M
Endo
Yaeta
Y
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000378:metabolism*; D002474:Cell-Free System; D016969:Desulfovibrio vulgaris / Q000378:metabolism*; D005486:Flavin Mononucleotide / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D013997:Time Factors
D001426:Bacterial Proteins; D005486:Flavin Mononucleotide