Rosado-Olivieri Edwin A, Ramos-Ortiz Gibram A, Hernández-Pasos Josué, Díaz-Balzac Carlos A, Vázquez-Rosa Edwin, Valentín-Tirado Griselle, Vega Irving E, García-Arrarás José E
Department of Biology, University of Puerto Rico-Río Piedras Campus, San Juan PR 00931, Puerto Rico.
Department of Biology, University of Puerto Rico-Río Piedras Campus, San Juan PR 00931, Puerto Rico.
Comp Biochem Physiol B Biochem Mol Biol. 2017 Dec;214:57-65. doi: 10.1016/j.cbpb.2017.08.004. Epub 2017 Aug 31.
One of the main challenges faced by investigators studying the nervous system of members of the phylum Echinodermata is the lack of markers to identify nerve cells and plexi. Previous studies have utilized an antibody, RN1, that labels most of the nervous system structures of the sea cucumber Holothuria glaberrima and other echinoderms. However, the antigen recognized by RN1 remained unknown. In the present work, the antigen has been characterized by immunoprecipitation, tandem mass spectrometry, and cDNA cloning. The RN1 antigen contains a START lipid-binding domain found in Steroidogenic Acute Regulatory (StAR) proteins and other lipid-binding proteins. Phylogenetic tree assembly showed that the START domain is highly conserved among echinoderms. We have named this antigen HgSTARD10 for its high sequence similarity to the vertebrate orthologs. Gene and protein expression analyses revealed an abundance of HgSTARD10 in most H. glaberrima tissues including radial nerve, intestine, muscle, esophagus, mesentery, hemal system, gonads and respiratory tree. Molecular cloning of HgSTARD10, consequent protein expression and polyclonal antibody production revealed the STARD10 ortholog as the antigen recognized by the RN1 antibody. Further characterization into this START domain-containing protein will provide important insights for the biochemistry, physiology and evolution of deuterostomes.
研究棘皮动物门成员神经系统的研究人员面临的主要挑战之一是缺乏用于识别神经细胞和神经丛的标记物。先前的研究使用了一种抗体RN1,它能标记海参光滑海参和其他棘皮动物的大部分神经系统结构。然而,RN1识别的抗原仍然未知。在目前的工作中,通过免疫沉淀、串联质谱和cDNA克隆对该抗原进行了表征。RN1抗原包含一个在类固醇生成急性调节(StAR)蛋白和其他脂质结合蛋白中发现的START脂质结合结构域。系统发育树组装显示,START结构域在棘皮动物中高度保守。由于其与脊椎动物直系同源物的高度序列相似性,我们将该抗原命名为HgSTARD10。基因和蛋白质表达分析显示,HgSTARD10在大多数光滑海参组织中大量存在,包括桡神经、肠道、肌肉、食道、肠系膜、血系统、性腺和呼吸树。HgSTARD10的分子克隆、随后的蛋白质表达和多克隆抗体制备揭示了STARD10直系同源物是RN1抗体识别的抗原。对这种含START结构域的蛋白质的进一步表征将为后口动物的生物化学、生理学和进化提供重要见解。