Longistatin, an EF-Hand Ca2+-Binding Protein from Vector Tick: Identification, Purification, and Characterization

Anisuzzaman, Anisuzzaman; Islam, M. Khyrul; Alim, Md. Abdul; Tsuji, Naotoshi

doi:10.1007/978-1-62703-230-8_9

Cited by 7 publications

(7 citation statements)

References 47 publications

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“…Similar to calreticulin, calmodulin is multifunctional protein that is primarily known for its roles in Ca 2+ homeostasis in mammals [168]. Except for a single study that described a H. longicornis tick calmodulin-like protein [169], nothing is known about this protein in tick physiology. Similarly, there are no studies on the roles of calponin in tick physiology.…”

Section: Resultsmentioning

confidence: 99%

A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome

et al. 2014

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BackgroundMultiple tick saliva proteins, the majority of which are unknown, confer tick resistance in repeatedly infested animals. The objective of this study was to identify the 24-48 h fed Amblyomma americanum tick saliva immuno-proteome. The 24-48 h tick-feeding phase is critical to tick parasitism as it precedes important events in tick biology, blood meal feeding and disease agent transmission. Fed male, 24 and 96 h fed female phage display cDNA expression libraries were biopanned using rabbit antibodies to 24 and 48 h fed A. americanum female tick saliva proteins. Biopanned immuno-cDNA libraries were subjected to next generation sequencing, de novo assembly, and bioinformatic analysis.ResultsMore than 800 transcripts that code for 24-48 h fed A. americanum immuno-proteins are described. Of the 895 immuno-proteins, 52% (464/895) were provisionally identified based on matches in GenBank. Of these, ~19% (86/464) show high level of identity to other tick hypothetical proteins, and the rest include putative proteases (serine, cysteine, leukotriene A-4 hydrolase, carboxypeptidases, and metalloproteases), protease inhibitors (serine and cysteine protease inhibitors, tick carboxypeptidase inhibitor), and transporters and/or ligand binding proteins (histamine binding/lipocalin, fatty acid binding, calreticulin, hemelipoprotein, IgG binding protein, ferritin, insulin-like growth factor binding proteins, and evasin). Others include enzymes (glutathione transferase, cytochrome oxidase, protein disulfide isomerase), ribosomal proteins, and those of miscellaneous functions (histamine release factor, selenoproteins, tetraspanin, defensin, heat shock proteins).ConclusionsData here demonstrate that A. americanum secretes a complex cocktail of immunogenic tick saliva proteins during the first 24-48 h of feeding. Of significance, previously validated immunogenic tick saliva proteins including AV422 protein, calreticulin, histamine release factor, histamine binding/lipocalins, selenoproteins, and paramyosin were identified in this screen, supporting the specificity of the approach in this study. While descriptive, this study opens opportunities for in-depth tick feeding physiology studies.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-518) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome

et al. 2014

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“…His-tagged longistatin (10 μg) was trapped with Talon Metal Affinity Resin (Clontech Laboratories Inc.) as described (53) and incubated with the membrane fraction (100 μg) of stimulated HUVECs in buffer A. After washing, proteins were eluted and separated by 12.5% SDS-PAGE under reducing conditions and transferred onto a nitrocellulose membrane.…”

Section: Discussionmentioning

confidence: 99%

Longistatin in tick saliva blocks advanced glycation end-product receptor activation

Anisuzzaman¹,

Hatta²,

Miyoshi³

et al. 2014

J. Clin. Invest.

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“…The membrane was then washed with washing buffer (10 mM Tris-HCl, 60 mM KCl, 5 mM MgCl 2 , pH 7.5) 3 times and then stained with ruthenium red staining solution for 20 min at room temperature. The ruthenium red staining solution was prepared by dissolving ruthenium red in the washing buffer to a final working concentration of 25 mg/L [25].…”

Section: Ruthenium Red Assay Of ∆Ltrinmentioning

confidence: 99%

“…The procedure for conducting the gel mobility assay was performed following the method described elsewhere [25]. Briefly, we prepared 15% SDS gel with 3 mM Ca 2+ and 3 mM EDTA.…”

Section: Gel Mobility Shift Assay Of ∆Ltrinmentioning

confidence: 99%

Recognition of Aedes aegypti Mosquito Saliva Protein LTRIN by the Human Receptor LTβR for Controlling the Immune Response

Loh,

Anthony,

Gavor

et al. 2024

Biology

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Salivary proteins from mosquitoes have received significant attention lately due to their potential to develop therapeutic treatments or vaccines for mosquito-borne diseases. Here, we report the characterization of LTRIN (lymphotoxin beta receptor inhibitor), a salivary protein known to enhance the pathogenicity of ZIKV by interrupting the LTβR-initiated NF-κB signaling pathway and, therefore, diminish the immune responses. We demonstrated that the truncated C-terminal LTRIN (ΔLTRIN) is a dimeric protein with a stable alpha helix-dominant secondary structure, which possibly aids in withstanding the temperature fluctuations during blood-feeding events. ΔLTRIN possesses two Ca2+ binding EF-hand domains, with the second EF-hand motif playing a more significant role in interacting with LTβR. Additionally, we mapped the primary binding regions of ΔLTRIN on LTβR using hydrogen–deuterium exchange mass spectrometry (HDX-MS) and identified that 91QEKAHIAEHMDVPIDTSKMSEQELQFHY118 from the N-terminal of ΔLTRIN is the major interacting region. Together, our studies provide insight into the recognition of LTRIN by LTβR. This finding may aid in a future therapeutic and transmission-blocking vaccine development against ZIKV.

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Longistatin, an EF-Hand Ca2+-Binding Protein from Vector Tick: Identification, Purification, and Characterization

Cited by 7 publications

References 47 publications

A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome

A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome

Longistatin in tick saliva blocks advanced glycation end-product receptor activation

Recognition of Aedes aegypti Mosquito Saliva Protein LTRIN by the Human Receptor LTβR for Controlling the Immune Response

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