Kentrup H, Becker W, Heukelbach J, Wilmes A, Schürmann A, Huppertz C, Kainulainen H, Joost H G
Institut für Pharmakologie und Toxikologie, Rheinisch-Westfälische Technische Hochschule Aachen, D-52057 Aachen, Federal Republic of Germany.
J Biol Chem. 1996 Feb 16;271(7):3488-95. doi: 10.1074/jbc.271.7.3488.
The cDNA of a novel, ubiquitously expressed protein kinase (Dyrk) was cloned from a rat brain cDNA library. The deduced amino acid sequence (763 amino acids) contains a catalytic domain that is only distantly related to that of other mammalian protein kinases. Its closest relative is the protein kinase Mnb of Drosophila, which is presumably involved in postembryonic neurogenesis (85% identical amino acids within the catalytic domain). Outside the catalytic domain, the sequence comprises several striking structural features: a bipartite nuclear translocation signal, a tyrosine-rich hydrophilic motif flanking the nuclear localization signal, a PEST region, a repeat of 13 histidines, a repeat of 17 serine/threonine residues, and an alternatively spliced insertion of nine codons. A recombinant glutathione S-transferase-Dyrk fusion protein catalyzed autophosphorylation and histone phosphorylation on tyrosine and serine/threonine residues with an apparent Km of approximately 3.4 microM. Exchange of two tyrosine residues in the "activation loop" between subdomains VII and VIII for phenylalanine almost completely suppressed the activity and tyrosine autophosphorylation of Dyrk. Tyrosine autophosphorylation was also reduced by exchange of the tyrosine (Tyr-219) in a tyrosine phosphorylation consensus motif. The data suggest that Dyrk is a dual specificity protein kinase that is regulated by tyrosine phosphorylation in the activation loop and might be a component of a signaling pathway regulating nuclear functions.
从大鼠脑cDNA文库中克隆出一种新型的、在全身广泛表达的蛋白激酶(Dyrk)的cDNA。推导的氨基酸序列(763个氨基酸)包含一个催化结构域,该结构域与其他哺乳动物蛋白激酶的催化结构域仅有远缘关系。其最相近的同源物是果蝇的蛋白激酶Mnb,推测其参与胚胎后神经发生(催化结构域内氨基酸序列同源性为85%)。在催化结构域之外,该序列包含几个显著的结构特征:一个双分型核转运信号、核定位信号两侧富含酪氨酸的亲水性基序、一个PEST区域、13个组氨酸的重复序列、17个丝氨酸/苏氨酸残基的重复序列以及一个由9个密码子组成的可变剪接插入序列。重组谷胱甘肽S-转移酶-Dyrk融合蛋白催化酪氨酸和丝氨酸/苏氨酸残基的自身磷酸化以及组蛋白磷酸化,其表观Km约为3.4 microM。将亚结构域VII和VIII之间“激活环”中的两个酪氨酸残基替换为苯丙氨酸几乎完全抑制了Dyrk的活性和酪氨酸自身磷酸化。在酪氨酸磷酸化共有基序中,酪氨酸(Tyr-219)的替换也降低了酪氨酸自身磷酸化。这些数据表明,Dyrk是一种双特异性蛋白激酶,其活性受激活环中酪氨酸磷酸化的调节,可能是调节核功能的信号通路的一个组成部分。