Protein profiling of hemolymph in Haemaphysalis flava ticks

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

WITHDRAWN: Molecular characterization of enolase fromHaemaphysalis longicornistick

“…4). Hemolymph in engorged adult H. flava was discovered using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in an earlier investigation (Liu et al, 2022).…”

Comprehensive Proteomics Analysis of the Hemolymph Composition of Sugar-Fed Aedes aegypti Female and Male Mosquitoes

“…Proteases and protease inhibitors are involved in many important processes to arthropod physiology such as development, reproduction, immunity, digestion, and even in vector-host interaction, as observed for salivary protease inhibitors found in some hematophagous species. ,− The presence of protease inhibitors has been described in the hemolymph of many other arthropod species where they were shown to be involved in regulating hemolymph clotting, immune responses, and antimicrobial defenses. ,,,− ,− In A. aegypti, it has been already reported that bacterial challenges led to higher expression of different serpins .…”

Proteomic analysis of extracellular vesicles from tick hemolymph and uptake of extracellular vesicles by salivary glands and ovary cells

“…By searching the UniProt Oryctolagus cuniculus database, we identified a total of 13 and 16 host proteins from RhHEEVs and HaHEEVs (Additional file 15 : Table S11), respectively. Among these, alpha-globin 1, hemoglobin, albumin, alpha-2-macroglobulin and histone H2A have been previously identified in tick saliva and hemolymph proteomics [ 52 , 53 ]. The presence of host-derived proteins suggests that tick hemolymph EVs may be involved in the transport of host proteins.…”

“…This is consistent with previous proteomic research on tick saliva [ 80 ]. However, Liu et al found that hemolymph collected from engorged Haemaphysalis flava was enriched with host proteins [ 53 ]. How these host proteins are transported from the midgut to the hemolymph is unclear.…”