Abstract
Abstract
Background
Tick hemolymph bathes internal organs, acts as an exchange medium for nutrients and cellular metabolites, and offers protection against pathogens. Hemolymph is abundant in proteins. However, there has been limited integrated protein analysis in tick hemolymph thus far. Moreover, there are difficulties in differentiating tick-derived proteins from the host source. The aim of this study was to profile the tick/host protein components in the hemolymph of Haemaphysalis flava.
Methods
Hemolymph from adult engorged H. flava females was collected by leg amputation from the Erinaceus europaeus host. Hemolymph proteins were extracted by a filter-aided sample preparation protocol, digested by trypsin, and assayed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). MS raw data were searched against the UniProt Erinaceidae database and H. flava protein database for host- and tick-derived protein identification. Protein abundance was further quantified by intensity-based absolute quantification (iBAQ).
Results
Proteins extracted from hemolymph unevenly varied in size with intense bands between 100 and 130 kDa. In total, 312 proteins were identified in the present study. Therein 40 proteins were identified to be host-derived proteins, of which 18 were high-confidence proteins. Top 10 abundant host-derived proteins included hemoglobin subunit-α and subunit-β, albumin, serotransferrin-like, ubiquitin-like, haptoglobin, α-1-antitrypsin-like protein, histone H2B, apolipoprotein A-I, and C3-β. In contrast, 169 were high-confidence tick-derived proteins. These proteins were classified into six categories based on reported functions in ticks, i.e., enzymes, enzyme inhibitors, transporters, immune-related proteins, muscle proteins, and heat shock proteins. The abundance of Vg, microplusin and α-2-macroglobulin was the highest among tick-derived proteins as indicated by iBAQ.
Conclusions
Numerous tick- and host-derived proteins were identified in hemolymph. The protein profile of H. flava hemolymph revealed a sophisticated protein system in the physiological processes of anticoagulation, digestion of blood meal, and innate immunity. More investigations are needed to characterize tick-derived proteins in hemolymph.
Graphical Abstract
Funder
Foundation for Distinguished Young Talents in Higher Education of Guangdong
the National Science Foundation for Young Scientists of China
the Double first-class construction project of Hunan Agricultural University
the Hunan Province Technology Breakthrough Project of 2021 for the open competition mechanism to select the best candidates
Publisher
Springer Science and Business Media LLC
Subject
Infectious Diseases,Parasitology
Reference80 articles.
1. Sonenshine DE. Biology of ticks. Oxford: Oxford University; 1992.
2. Libor G, Nataliia R, Marie V, Maryna G, Jan S. Circulatory system and hemolymph. In: Sonenshine DE, Roe R, editors. Biology of ticks. Oxford: Oxford University; 2013. p. 263.
3. James AM, Oliver JH. Purification and partial characterization of vitellin from the black-legged tick, Ixodes scapularis. Insect Biochem Mol Biol. 1997;27:639–49.
4. Chinzei Y, Chino H, Takahashi K. Purification and properties of vitellogenin and vitellin from a tick, Ornithodoros moubata. J Comp Physiol. 1983;152:13–21.
5. Kopacek P, Weise C, Saravanan T, Vitova K, Grubhoffer L. Characterization of an αmacroglobulin-like glycoprotein isolated from the plasma of the soft tick Ornithodoros moubata. Eur J Biochem. 2000;267:465–75.