文章基本信息

标题：MECHANISM AND REGULATION OF THIAMINE PYROPHOSPHOKINASE FROM PARSELY LEAF
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作者：Hisateru MITSUDA ; Yukio TAKII ; Kimikazu IWAMI 等
期刊名称：Journal of Nutritional Science and Vitaminology
印刷版ISSN：0301-4800
电子版ISSN：1881-7742
出版年度：1975
卷号：21
期号：3
页码：189-198
DOI：10.3177/jnsv.21.189
出版社：Center for Academic Publications Japan
摘要：Thiamine pyrophosphokinase (EC 2.7.6.2) from parsely leaf showed an absolute requirement for divalent cation such as Mg²⁺, Mn²⁺ and Co²⁺. The activation effect varied with the species and concentrations of such cations. When Mn²⁺ or Co²⁺ was used as cofactor, maximal activation was found at a lower level than ATP concentration, whereas the activation by Mg²⁺ increased hyperbolically with the concentration. Studies of initial velocity and product inhibition led to conclude that the kinase reaction obeys a sequential ordered Bi Bi mechanism; i.e. the enzyme combines in turns with MgATP and thiamine, followed by release of TPP and AMP. The inhibition type revealed for inorganic pyrophosphate was competitive with respect to thiamine with K_i of approximately 2.8mM. On the other hand, thiamine monophosphate exhibited noncompetitive inhibition with K_i of 0.2mM. The plots of the reaction rate against MgATP concentrations gave a sigmoidal curve. Addition of either AMP or GMP resulted in restoration of a depressed activity at low concentration of MgATP. The “allosteric” inhibition was also relieved by the addition of an excess amount of magnesium ions. These findings suggest that transphosphorylation is regulated by subcellular concentrations of metal ions relative to ATP or of the products involved in the thiamine biosynthesis.