Singh Vijay, Nair Divya N, Kaushal Radhey Shyam, Kumar Manoj, Pappachan Anju, Singh Desh Deepak
Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India; Centre for Genetic Disease and Molecular Medicine, Central University of Punjab, Bathinda 151001, India.
Biotechnol Rep (Amst). 2015 Oct 3;8:81-87. doi: 10.1016/j.btre.2015.09.004. eCollection 2015 Dec.
Leishmaniasis, a disease of the developing world affects about 12 million people and has limited therapeutic interventions available. L-type lectins, Endoplasmic Reticulum Golgi Intermediate Compartment/Vesicular Integral Proteins (ERGIC-53/VIP36) are involved in protein sorting in luminal compartments of animal cells and are important for parasite biology. A lectin homologue was identified through a bioinformatics analysis of genome and it was found to have N-terminal conserved carbohydrate recognition domain (CRD) and a unique C-terminal region rich in repetitive amino acids and a poly glutamine tract. The N-terminal CRD region was cloned and expressed in , but gave an insoluble expression which was re-solubilized by on column refolding. The fold integrity was checked through CD, fluorescence and functional assay of hemagglutination activity using rabbit erythrocyte. Bioinformatics analysis identified 15 members from Tritryps ( spp., spp.) and they separate out as a distinct clade in the global phylogenetic analysis of all ERGIC-53/VIP36 sequences downloaded from Uniprot. Our analysis shows that the extended C-terminal regions with repeats is unique to Tritryps and this repeat pattern is different in sequences from spp and spp and all these features make this protein an interesting candidate for further detailed studies.
利什曼病是一种在发展中国家流行的疾病，影响着约1200万人，且可用的治疗干预措施有限。L型凝集素，即内质网高尔基体中间腔/囊泡整合蛋白（ERGIC-53/VIP36），参与动物细胞腔内区室的蛋白质分选，对寄生虫生物学具有重要意义。通过对基因组进行生物信息学分析鉴定出一种凝集素同源物，发现它具有N端保守的碳水化合物识别结构域（CRD）以及富含重复氨基酸和多聚谷氨酰胺序列的独特C端区域。N端CRD区域被克隆并在 中表达，但表达产物不溶，通过柱上重折叠使其重新溶解。通过圆二色光谱（CD）、荧光以及使用兔红细胞进行血凝活性的功能测定来检查折叠完整性。生物信息学分析在三锥虫属（ 种、 种）中鉴定出15个成员，在从Uniprot下载的所有ERGIC-53/VIP36序列的全球系统发育分析中，它们作为一个独特的进化枝分离出来。我们的分析表明，具有重复序列的扩展C端区域是三锥虫属所特有的，并且这种重复模式在 种和 种的序列中有所不同，所有这些特征使得这种蛋白质成为进一步详细研究的有趣候选对象。
