Parvalbumins play crucial physiological roles in neuromuscular systems of vertebrates, such as cell-cycle, development of neurons, contraction of muscles, and regulation of intracellular calcium. To perform these neuromuscular functions, parvalbumin may be in associated with other proteins including calbindin, carbonic anhydrase, and cytochrome oxidase. Humans may show an IgE-specific hypersensitivity to parvalbumins after consumption of some distinct fish species. While this protein is abundant in fish muscles, literature review of publications related to fish parvalbumins, do not point to the presence of parvalbumins in eukaryotic microbes. In this study, we propose that distantly related parvalbumins may be found in some non-fish species. Bioinformatics studies such as multiple sequence alignment (MSA), phylogenetic analysis as well as molecular-based experiments indicate that, at least two parvalbumins sequences (UniProt IDs: A0A178F775 and A0A178F7E4) with EF-hand domains and Ca2+-binding sites could be identified in Trichophyton violaceum, a pathogenic fungal species. It was determined that both genes consisted of a single exon and encoded for parvalbumin proteins possessing conserved amino acid motifs. Antigenicity prediction revealed antigenic sites located in both sides of the Ca2+-binding site of the first EF-hand domain. Our phylogenetic analysis revealed that one of parvalbumins (UniProt ID: 0A178F775) can be evolved to other parvalbumins in T. violaceum (UniProt ID: A0A178F7E4) and fish species through evolutionary phenomenon. To confirm our in-silico findings, we designed three primer pairs to detect one of the T. violaceum parvalbumins (UniProt ID: A0A178F7E4) by polymerase chain reaction (PCR); one primer pair showed a strong and specific band in agarose gel electrophoresis. To evaluate the specificity of the method, the primers were tested on extracted DNA from Trichophyton rubrum and T. mentagrophytes. The results demonstrated that the evaluated parvalbumin gene (UniProt ID: A0A178F7E4) was T. violaceum-specific and this pathogenic fungus can be differentiated from T. rubrum and T. mentagrophytes through identification of parvalbumin genes. Further studies are necessary to unravel the biochemical and physiological functions of parvalbumins in T. violaceum.

中文翻译：

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紫色毛癣菌中小清蛋白样蛋白的鉴定和表征

小清蛋白在脊椎动物的神经肌肉系统中起着至关重要的生理作用，例如细胞周期、神经元发育、肌肉收缩和细胞内钙的调节。为了执行这些神经肌肉功能，小清蛋白可能与其他蛋白质相关，包括钙结合蛋白、碳酸酐酶和细胞色素氧化酶。人类在食用某些不同的鱼类后可能会表现出对小清蛋白的 IgE 特异性超敏反应。虽然这种蛋白质在鱼类肌肉中含量丰富，但与鱼类小清蛋白相关的出版物的文献综述并未指出真核微生物中存在小清蛋白。在这项研究中，我们提出可能在一些非鱼类物种中发现远亲小清蛋白。生物信息学研究，例如多序列比对 (MSA)，系统发育分析以及基于分子的实验表明，至少可以在紫色毛癣菌（一种病原真菌）中鉴定出两个具有 EF 手结构域和 Ca2+ 结合位点的小清蛋白序列（UniProt ID：A0A178F775 和 A0A178F7E4）。确定两个基因均由单个外显子组成并编码具有保守氨基酸基序的小清蛋白蛋白质。抗原性预测显示抗原位点位于第一个 EF 手结构域的 Ca2+ 结合位点两侧。我们的系统发育分析表明，一种小白蛋白（UniProt ID：0A178F775）可以​​通过进化现象进化为紫罗兰（UniProt ID：A0A178F7E4）和鱼类中的其他小白蛋白。为了证实我们的计算机研究结果，我们设计了三个引物对来检测 T 中的一个。violaceum parvalbumins (UniProt ID: A0A178F7E4) 通过聚合酶链反应 (PCR)；一对引物在琼脂糖凝胶电泳中显示出强而特异的条带。为了评估该方法的特异性，在从红色毛癣菌和须须癣菌中提取的 DNA 上测试了引物。结果表明，所评估的小白蛋白基因（UniProt ID：A0A178F7E4）是紫色毛癣菌特异性的，通过鉴定小白蛋白基因可以将这种病原真菌与红色毛癣菌和须癣毛癣菌区分开来。需要进一步的研究来阐明 T. violaceum 中小清蛋白的生化和生理功能。引物在从红色毛癣菌和须癣毛癣菌中提取的 DNA 上进行测试。结果表明，所评估的小白蛋白基因（UniProt ID：A0A178F7E4）是紫色毛癣菌特异性的，通过鉴定小白蛋白基因可以将这种病原真菌与红色毛癣菌和须癣毛癣菌区分开来。需要进一步的研究来阐明 T. violaceum 中小清蛋白的生化和生理功能。引物在从红色毛癣菌和须癣毛癣菌中提取的 DNA 上进行测试。结果表明，所评估的小白蛋白基因（UniProt ID：A0A178F7E4）是紫色毛癣菌特异性的，通过鉴定小白蛋白基因可以将这种病原真菌与红色毛癣菌和须癣毛癣菌区分开来。需要进一步的研究来阐明 T. violaceum 中小清蛋白的生化和生理功能。