Dea-Ayuela María Auxiliadora, Rama-Iñiguez Sara, Bolás-Fernández Francisco
Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense, Madrid, Spain.
Proteomics. 2006 Jul;6(14):4187-94. doi: 10.1002/pmic.200600101.
Leishmaniasis is a zoonotic disease caused by the species of the genus Leishmania, flagellated protozoa that multiply inside mammalian macrophages and are transmitted by the bite of the sandfly. The disease is widespread and due to the lack of fully effective treatment and vaccination the search for new drugs and immune targets is needed. Proteomics seems to be a suitable strategy because the annotated sequenced genome of L. major is available. Here, we present a high-resolution proteome for L. infantum promastigotes comprising of around 700 spots. Western blot with rabbit hyperimmune serum raised against L. infantum promastiogote extracts and further analysis by MALDI-TOF and MALDI-TOF/TOF MS allowed the identification of various relevant functional antigenic proteins. Major antigenic proteins were identified as propionil carboxilasa, ATPase beta subunit, transketolase, proteasome subunit, succinyl-diaminopimelate desuccinylase, a probable tubulin alpha chain, the full-size heat shock protein 70, and several proteins of unknown function. In addition, one enzyme from the ergosterol biosynthesis pathway (adrenodoxin reductase) and the structural paraflagellar rod protein 3 (PAR3) were found among non-antigenic proteins. This study corroborates the usefulness of proteomics in identifying new proteins with crucial biological functions in Leishmania parasites.
利什曼病是一种人畜共患疾病，由利什曼原虫属的物种引起，这些有鞭毛的原生动物在哺乳动物巨噬细胞内繁殖，并通过白蛉叮咬传播。该疾病广泛传播，由于缺乏完全有效的治疗方法和疫苗，需要寻找新的药物和免疫靶点。蛋白质组学似乎是一种合适的策略，因为硕大利什曼原虫的注释测序基因组是可用的。在这里，我们展示了婴儿利什曼原虫前鞭毛体的高分辨率蛋白质组，包含约700个斑点。用针对婴儿利什曼原虫前鞭毛体提取物产生的兔超免疫血清进行蛋白质免疫印迹，并通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）和基质辅助激光解吸电离串联飞行时间质谱（MALDI-TOF/TOF MS）进一步分析，从而鉴定出各种相关的功能性抗原蛋白。主要抗原蛋白被鉴定为丙酰羧化酶、ATP酶β亚基、转酮醇酶、蛋白酶体亚基、琥珀酰二氨基庚二酸脱琥珀酰酶、一种可能的微管蛋白α链、全长热休克蛋白70以及几种功能未知的蛋白质。此外，在非抗原蛋白中发现了一种来自麦角固醇生物合成途径的酶（肾上腺皮质铁氧化还原蛋白还原酶）和结构副鞭毛杆蛋白3（PAR3）。这项研究证实了蛋白质组学在鉴定利什曼原虫中具有关键生物学功能的新蛋白质方面的有用性。
