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松材线虫(Bursaphelenchus xylophilus)的三种乙酰胆碱酯酶:对不同生理功能的见解

Three acetylcholinesterases of the pinewood nematode, Bursaphelenchus xylophilus: insights into distinct physiological functions.

作者信息

Kang Jae Soon, Lee Dae-Weon, Choi Jae Young, Je Yeon Ho, Koh Young Ho, Lee Si Hyeock

机构信息

Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea.

出版信息

Mol Biochem Parasitol. 2011 Feb;175(2):154-61. doi: 10.1016/j.molbiopara.2010.11.005. Epub 2010 Nov 11.

Abstract

Acetylcholinesterase (AChE) plays a key role in postsynaptic transmission in most animals. Nematodes encode multiple AChEs, implying its functional diversity. To explore physiological functions of multiple AChEs, three distinct AChEs (BxACE-1, BxACE-2, and BxACE-3) were identified and characterized from the pinewood nematode. Sequencing comparison with Torpedo AChE and Caenorhabditis elegans ACEs identified choline-binding site, catalytic triad functional site, three internal disulfide bonds and aromatic residues for the catalytic gorge. Transcriptional profiling by quantitative real-time PCR revealed that BxACE-3 is more actively transcribed than BxACE-1 (2-3 times) and BxACE-2 (9-18 times) in both propagative and dispersal stages. The three BxACEs were functionally expressed using baculovirus system. Kinetic analysis of in vitro-expressed BxACEs revealed that the substrate specificity was highest in BxACE-1 whereas the catalytic efficiency was highest in BxACE-2. In inhibition assay, BxACE-3 showed the lowest inhibition rate. Taken together, it appears that both BxACE-1 and BxACE-2 play common but non-overlapping roles in synaptic transmission, whereas BxACE-3 may have non-neuronal functions. The current findings should provide valuable insights into the evolutionary process and various physiological roles of AChE.

摘要

乙酰胆碱酯酶(AChE)在大多数动物的突触后传递中起关键作用。线虫编码多种AChE,这意味着其功能具有多样性。为了探究多种AChE的生理功能,从松材线虫中鉴定并表征了三种不同的AChE(BxACE - 1、BxACE - 2和BxACE - 3)。与电鳗AChE和秀丽隐杆线虫ACEs的序列比较确定了胆碱结合位点、催化三联体功能位点、三个内部二硫键以及催化峡谷的芳香族残基。通过定量实时PCR进行的转录谱分析表明,在繁殖期和扩散期,BxACE - 3的转录活性均高于BxACE - 1(2 - 3倍)和BxACE - 2(9 - 18倍)。利用杆状病毒系统对三种BxACEs进行了功能表达。对体外表达的BxACEs的动力学分析表明,BxACE - 1的底物特异性最高,而BxACE - 2的催化效率最高。在抑制试验中,BxACE - 3的抑制率最低。综上所述,BxACE - 1和BxACE - 2似乎在突触传递中发挥共同但不重叠的作用,而BxACE - 3可能具有非神经元功能。目前的研究结果应为AChE的进化过程和各种生理作用提供有价值的见解。

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