Janowitz Tim, Kneifel Helmut, Piotrowski Markus
Department of Plant Physiology, Ruhr-Universität, Universitätsstr. 150, D-44801 Bochum, Germany.
FEBS Lett. 2003 Jun 5;544(1-3):258-61. doi: 10.1016/s0014-5793(03)00515-5.
The cloning, expression and characterization of plant agmatine iminohydrolase (AIH, also known as agmatine deiminase, EC 3.5.3.12) is described. Recombinant AIH of Arabidopsis thaliana forms dimers and catalyzes the specific conversion of agmatine to N-carbamoylputrescine and ammonia. Biochemical data suggested that cysteine side chains are involved in catalysis. However, site-directed mutagenesis of the two highly conserved cysteine residues of AIH showed that these cysteines are important but not essential for activity, arguing against a thioester substrate-enzyme intermediate during catalysis. This work represents the completion of the cloning of the arginine decarboxylase pathway genes of higher plants.
本文描述了植物胍丁胺亚胺水解酶(AIH,也称为胍丁胺脱亚氨酶,EC 3.5.3.12)的克隆、表达及特性。拟南芥重组AIH形成二聚体,并催化胍丁胺特异性转化为N-氨甲酰腐胺和氨。生化数据表明,半胱氨酸侧链参与催化过程。然而,对AIH两个高度保守的半胱氨酸残基进行定点诱变后发现,这些半胱氨酸对活性很重要,但并非必需,这与催化过程中硫酯底物-酶中间体的观点相悖。这项工作代表了高等植物精氨酸脱羧酶途径基因克隆的完成。