CSIRO Livestock and Aquaculture, Queensland Bioscience Precinct, 306 Carmody Rd, St Lucia, QLD, Australia.
James Cook University, Molecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook Drive, Townsville, QLD 4811, Australia; James Cook University, Australian Institute of Tropical Health and Medicine, James Cook Drive, Townsville, QLD 4811, Australia.
J Proteomics. 2020 Apr 30;218:103689. doi: 10.1016/j.jprot.2020.103689. Epub 2020 Feb 21.
Hemocyanin (Hc) is a multifunctional macromolecule involved in oxygen transport and non-specific immunity in shrimp. Hc is crucial in physiology and nutrition linked with optimal performance in aquaculture production systems. In medicine, Hc has been approved for clinical use in humans as adjuvant and anticancer therapeutic. In contrast, Hc has also been identified as one of the proteins causing anaphylaxis following shrimp consumption. The role of individual Hc isoforms remains unknown due to a lack of resolved Hc isoforms. We successfully identified eleven different Penaeus monodon hemocyanin (PmoHc) γ isoforms including two truncated isoforms (50 and 20 kDa) and one PmoHc β isoform in haemolymph using proteomics informed by transcriptomics. Amino acid sequence homology ranged from 24 to 97% between putative PmoHc gene isoforms. Hc isoforms showed specific patterns of transcript expression in shrimp larval stages and adult hepatopancreas. These findings enable isoform level investigations aiming to define molecular mechanisms underpinning Hc functionality in shrimp physiology and immunity, as well as their individual immunogenic role in human allergy. Our research demonstrates the power of proteomics informed by transcriptomics to resolve isoform complexity in non-model organisms and lay the foundations for improved performance within the aquaculture industry and advance allergenic applications in medicine. SIGNIFICANCE: The roles of hemocyanin (Hc) in shrimp homeostasis and immunity as well as in human allergy are not well understood because the complexity of Hc isoforms has remained unresolved. Our results have confirmed the existence of at least 12 individual Hc isoforms in shrimp haemolymph and validated putative Hc gene assemblies from transcriptomics. Our findings will enable monitoring the expression of specific Hc isoforms in shrimp haemolymph during different environmental, nutritional and pathogenic conditions, thus providing insights into isoform specific functional roles. In medicine, the potential allergenicity of each Hc isoform could be determined and advance allergenic applications. Lastly, since Hc comprises up to 95% of the total protein in haemolymph, these isoforms become ideal targets for prawn provenance, traceability and food contamination studies.
血蓝蛋白（Hc）是一种多功能大分子，参与虾类的氧气运输和非特异性免疫。Hc 在与水产养殖生产系统中最佳性能相关的生理学和营养方面至关重要。在医学上，Hc 已被批准在人类中作为佐剂和抗癌治疗药物使用。相比之下，Hc 也被确定为虾类消费后引起过敏的蛋白质之一。由于缺乏已解决的 Hc 同工型，个体 Hc 同工型的作用仍然未知。我们使用基于转录组学的蛋白质组学成功鉴定了 11 种不同的斑节对虾血蓝蛋白（PmoHc）γ同工型，包括两种截断同工型（50 和 20 kDa）和一种 PmoHcβ同工型。假定的 PmoHc 基因同工型之间的氨基酸序列同源性范围为 24%至 97%。Hc 同工型在虾幼体和成年肝胰腺阶段表现出特定的转录表达模式。这些发现使同工型水平的研究能够定义 Hc 功能在虾生理学和免疫中的分子机制，以及它们在人类过敏中的个体免疫作用成为可能。我们的研究表明，基于转录组学的蛋白质组学能够解决非模式生物中的同工型复杂性，并为水产养殖行业的性能提升和医学中变应原应用的发展奠定基础。
血蓝蛋白（Hc）在虾类体内平衡和免疫中的作用以及在人类过敏中的作用尚不清楚，因为 Hc 同工型的复杂性尚未得到解决。我们的结果证实了虾血淋巴中至少存在 12 种个体 Hc 同工型，并验证了来自转录组学的假定 Hc 基因组装。我们的发现将能够在不同的环境、营养和病理条件下监测虾血淋巴中特定 Hc 同工型的表达，从而深入了解同工型特异性功能作用。在医学上，可以确定每个 Hc 同工型的潜在变应原性，并推进变应原应用。最后，由于 Hc 占血淋巴总蛋白的 95%，因此这些同工型成为对虾产地、可追溯性和食物污染研究的理想目标。
