Abstract：
Fibrillin is an important structural protein in connective tissues. The presence of fibrillin in sea cucumber Apostichopus japonicus is still poorly understood, which limits our understanding of the role of fibrillin in the A. japonicus microstructure. The aim of this study was to clarify the presence of fibrillin in the sea cucumber A. japonicus body wall. Herein, the presence of fibrillin in sea cucumber A. japonicus was investigated by utilizing targeted proteomics and visualization strategies. The contents of three different isoforms of fibrillin with high abundance in A. japonicus were determined to be 0.96, 2.54, and 0.15 μg/g (wet base), respectively. The amino acid sequence of fibrillin (GeneBank number: PIK56741.1) that started at position 631 and ended at position 921 was selected for cloning and expressing antigen. An anti-A. japonicus fibrillin antibody with a titer greater than 1:64 000 was successfully obtained. It was observed that the distribution of fibrillin in the A. japonicus body wall was scattered and dispersed in the form of fibril bundles at the microscale. It further observed that fibrillin was present near collagen fibrils and some entangled outside the collagen fibrils at the nanoscale. Moreover, the stoichiometry of the most dominant collagen and fibrillin molecules in A. japonicus was determined to be approximately 250:1. These results contribute to an understanding of the role of fibrillin in the sea cucumber microstructure.
摘要：
纤溶蛋白是结缔组织中重要的结构蛋白。海参刺参中原纤维蛋白的存在仍然知之甚少，这限制了我们对原纤蛋白在刺槐微观结构中的作用的理解。本研究的目的是阐明原纤蛋白在海参A体壁中的存在。在这里，利用靶向蛋白质组学和可视化策略研究了海参A中原纤维蛋白的存在。测定了刺槐中高丰度的三种不同原纤亚型的含量分别为0.96、2.54和0.15μg/g（湿基），分别。选择在631位开始并在921位结束的原纤维蛋白的氨基酸序列(GeneBank编号:PIK56741.1)用于克隆和表达抗原。反A。成功获得效价大于1:64000的刺血原纤维蛋白抗体。观察到原纤蛋白在刺槐体壁中的分布在微观尺度上以原纤束的形式分散和分散。进一步观察到纤丝蛋白存在于胶原纤维附近,并且一些在纳米级的胶原纤维外部缠结。此外，在刺槐中最主要的胶原和纤丝蛋白分子的化学计量被确定为大约250:1。这些结果有助于理解原纤蛋白在海参微观结构中的作用。
