Bouvier Leon A, Miranda Mariana R, Canepa Gaspar E, Alves Maria Júlia M, Pereira Claudio A
Departamento de Sustancias Vasoactivas, Instituto de Investigaciones Médicas Alfredo Lanari, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Combatientes de Malvinas, Buenos Aires, Argentina.
Biochim Biophys Acta. 2006 Jun;1760(6):913-21. doi: 10.1016/j.bbagen.2006.02.013. Epub 2006 Mar 13.
Adenylate kinases supply energy routes in cellular energetic homeostasis. In this work, we identified and characterized the adenylate kinase activity in extracts from the flagellated parasite Trypanosoma cruzi, the causative agent of Chagas' disease. Adenylate kinase activity was detected in different subcellular fractions and the cytosolic isoform was biochemically characterized. Cytosolic adenylate kinase specific activity increases continuously during the epimastigote growth and is down-regulated when other soluble phosphotransferase, arginine kinase, is overexpressed. Six different genes of adenylate kinase isoforms were identified and the mRNA expression was confirmed by RT-PCR and Northern Blot. Three open reading frames coding for different enzyme isoforms named TzADK1, TzADK2 and TzADK5 were cloned and functionally expressed in E. coli. This work reports an unusually large number of genes of adenylate kinases and suggests a coordinated regulation of phosphotransferase-mediated ATP regenerating pathways in the unicellular parasite Trypanosoma cruzi.
腺苷酸激酶在细胞能量稳态中提供能量途径。在这项研究中,我们鉴定并表征了来自鞭毛虫寄生虫克氏锥虫(恰加斯病的病原体)提取物中的腺苷酸激酶活性。在不同的亚细胞组分中检测到了腺苷酸激酶活性,并对胞质同工型进行了生化表征。胞质腺苷酸激酶的比活性在无鞭毛体生长过程中持续增加,而当其他可溶性磷酸转移酶精氨酸激酶过表达时则下调。鉴定出了六个不同的腺苷酸激酶同工型基因,并通过逆转录聚合酶链反应(RT-PCR)和Northern印迹法证实了其mRNA表达。克隆了编码不同酶同工型的三个开放阅读框,分别命名为TzADK1、TzADK2和TzADK5,并在大肠杆菌中进行了功能表达。这项研究报道了数量异常多的腺苷酸激酶基因,并表明单细胞寄生虫克氏锥虫中磷酸转移酶介导的ATP再生途径存在协同调节。