Different forms of molybdenum cofactor inVicia faba seeds: The presence of molybdenum cofactor carrier protein and its purification |
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Authors: | Kyrill L Kalakoutskii Emilio Fernândez |
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Institution: | (1) Departamento de Bioquimica y Biologia Molecular, Avda. San Alberto Magno s/n, Facultad de Ciencias, Universidad de Côrdoba, E-14071 Côrdoba, Spain;(2) Present address: A.N. Bach Institute of Biochemistry, Russian Academy of Scieneies, Leninsky prosp. 33, 117071 Moscow, Russia |
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Abstract: | There were significant differences in the contents of molybdenum cofactor (Mo-co), both in a low-molecular-mass form (free Mo-co) and in a protein-bound form, in seeds of sevenVicia faba genotypes. Low-molecular-mass Mo-co species present in the extracts were detected by their ability to reactivate, through a dialysis membrane, aponitrate reductase from theNeurospora crassa nit-1 mutant. In extracts of all genotypes tested, the amount of Mo-co capable of directly reactivating nitrate reductase of theN. crassa nit-1 mutant was always much higher than that of low-molecular-mass Moco. These data cannot be explained by considering, as traditionally, that Mo-co detected directly, i.e. without any previous treatment for its release from Mo-coproteins, corresponds to free low-molecular mass Mo-co. A protein which bound Mo-co was purified to electrophoretic homogeneity. This protein consisted of a single 70-kDa polypeptide chain and carried a Mo-co that could be efficiently released when in contact with aponitrate reductase.Abbreviations CP
carrier protein
- Mo-co
molybdenum cofactor
- NR
nitrate reductase
- XO
xanthine oxidase |
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Keywords: | Molybdate Molybdenum cofactor Nitrate reductase Vicia |
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