来自黎凡特蝰蛇毒液的5'-核苷酸酶。
5'-Nucleotidase from Vipera lebetina venom.
作者信息
Trummal Katrin, Samel Mari, Aaspõllu Anu, Tõnismägi Külli, Titma Tiina, Subbi Juhan, Siigur Jüri, Siigur Ene
机构信息
National Institute of Chemical Physics and Biophysics, Tallinn, Estonia.
National Institute of Chemical Physics and Biophysics, Tallinn, Estonia; Tallinn University, Tallinn, Estonia.
出版信息
Toxicon. 2015 Jan;93:155-63. doi: 10.1016/j.toxicon.2014.11.234. Epub 2014 Nov 28.
5'-Nucleotidase (5'-NT) is widely represented in animal tissues (CD73) as well as in almost all snake venoms. In the present study, a 5'-NT isoform has been isolated from Vipera lebetina venom. The homodimeric isoform consists of monomers with molecular masses of 60 kDa. The enzyme is thermolabile and has pH optimum at 7.5. The 5'-NT activity is inhibited by metal ions Fe(3+), Cu(2+) and Zn(2+), enhanced by Mn(2+) while Mg(2+) and Ca(2+) have no remarkable effect. In addition to 120-kDa protein there are higher molecular forms of 5'-NT present in the V. lebetina venom. The cloning and sequencing of the 5'-NT coding cDNA resulted in 5'-truncated construct. MALDI-TOF and Orbitrap mass-spectrometry of the tryptic peptides confirmed the translated N-terminally truncated protein sequence concordance to the 5'-NT isolated from the venom. The isolated protein strongly inhibited ADP- or collagen-induced platelet aggregation.
5'-核苷酸酶（5'-NT）在动物组织（CD73）以及几乎所有蛇毒中广泛存在。在本研究中，从黎凡特蝰蛇毒中分离出了一种5'-NT同工型。该同二聚体同工型由分子量为60 kDa的单体组成。该酶对热不稳定，最适pH为7.5。5'-NT活性受到金属离子Fe(3+)、Cu(2+)和Zn(2+)的抑制，被Mn(2+)增强，而Mg(2+)和Ca(2+)没有显著影响。除了120 kDa的蛋白质外，黎凡特蝰蛇毒中还存在更高分子形式的5'-NT。5'-NT编码cDNA的克隆和测序产生了5'-截短构建体。胰蛋白酶肽段的基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）和轨道阱质谱证实了翻译后的N端截短蛋白序列与从毒液中分离出的5'-NT一致。分离出的蛋白质强烈抑制ADP或胶原诱导的血小板聚集。
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