Pérez-Castiñeira José R, Alvar Jorge, Ruiz-Pérez Luis M, Serrano Aurelio
Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-CSIC, Avda. Americo Vespucio s/n, Sevilla 41092, Spain.
Biochem Biophys Res Commun. 2002 Jun 14;294(3):567-73. doi: 10.1016/S0006-291X(02)00517-X.
Proton-translocating inorganic pyrophosphatases (H(+)-PPase, EC 3.6.1.1) are integral membrane proteins that have been extensively studied in higher plants, the photosynthetic bacterium Rhodospirillum rubrum and, more recently, in some human pathogenic protozoa. By using a PCR-based approach, fragments of genes coding for H(+)-PPases in a number of protists, both free-living and parasites of animals and plants, that belong to diverse taxonomic groups (trypanosomatids, ciliates, apicomplexans, euglenoids, amoeboid mycetozoa, heterokonts) have been isolated. The experimental procedure involved the use of degenerate oligonucleotides designed from protein domains conserved in H(+)-PPases from plants and bacteria. The PCR-amplified DNA fragments exhibited the characteristic genomic structure and codon usage of the corresponding protozoan group. Paralogous genes were found in some species suggesting the occurrence of protein isoforms. These results indicate that H(+)-PPases are more widely distributed among protozoa than previously thought.
质子转运无机焦磷酸酶(H(+)-PPase,EC 3.6.1.1)是整合膜蛋白,已在高等植物、光合细菌红螺菌中得到广泛研究,最近在一些人类致病原生动物中也有研究。通过基于PCR的方法,已分离出多种原生生物中编码H(+)-PPase的基因片段,这些原生生物包括自由生活的以及动植物寄生虫,它们属于不同的分类群(锥虫、纤毛虫、顶复体虫、裸藻、变形菌菌绒孢菌、异鞭毛类)。实验过程涉及使用根据植物和细菌中H(+)-PPase保守蛋白结构域设计的简并寡核苷酸。PCR扩增的DNA片段表现出相应原生动物类群的特征性基因组结构和密码子使用情况。在一些物种中发现了旁系同源基因,表明存在蛋白质异构体。这些结果表明,H(+)-PPase在原生动物中的分布比以前认为的更广泛。