Characterization of glutamine: fructose-6-phosphate aminotransferase from the ixodid tick, Haemaphysalis longicornis, and its critical role in host blood feeding

A cDNA encoding the vitellogenin receptor of the ixodid tick, Haemaphysalis longicornis Neumann (HlVgR) was cloned and characterized. The full-length cDNA is 5631 bp, including an intact ORF encoding an expected protein with 1782 amino acids. The deduced amino acid sequence of the HlVgR cDNA revealed two ligand-binding domains with four class A cysteine-rich repeats in the first domain and eight in the second domain similar to those of insect VgRs. The immunoblot analysis detected ~197 kDa protein in both tick ovary and egg. The developmental expression profile demonstrated that HlVgR mRNA exists throughout the ovarian development, and the transcriptional level is especially high in the previtellogenic period. Immuno electron microscopy analysis demonstrated that the localization of HlVgR is detected on the external surface of oocyte plasma membrane. RNAi showed that eggs of HlVgR dsRNA-injected adult ticks had not developed into fully mature oocytes and laid abnormal eggs. The Babesia parasite DNA was not detected in the eggs of HlVgR dsRNA-injected tick that fed on Babesia gibsoni infected dog, whereas it was detected in the eggs of PBS-injected ticks and noninjected ticks. Expression of HlVgR was increased by the vitellogenic hormone 20-hydroxyecdysone. These results indicate that HlVgR, which is produced by the developing oocytes, is essential for Vg uptake, egg development in the H. longicornis tick, and transovarial transmission of Babesia parasites.

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Valosin-containing protein from the hard tick Haemaphysalis longicornis: effects of dsRNA-mediated HlVCP gene silencing

Biochemistry and Cell Biology ◽

10.1139/o07-051 ◽

2007 ◽

Vol 85 (3) ◽

pp. 384-394 ◽

Cited By ~ 8

Author(s):

Damdinsuren Boldbaatar ◽

Badgar Battsetseg ◽

Takeshi Hatta ◽

Takeharu Miyoshi ◽

Naotoshi Tsuji ◽

...

Keyword(s):

Blood Feeding ◽

Pcr Analysis ◽

Hard Tick ◽

Haemaphysalis Longicornis ◽

Native Protein ◽

Rt Pcr ◽

Future Studies ◽

Atpase Gene ◽

A Domains

We report the cloning and characterization of a cDNA encoding the valosin-containing protein (VCP) from the Haemaphysalis longicornis tick (HlVCP). The full-length HlVCP is 2782 bp and codes for 808 amino acids of a deduced protein with a predicted molecular mass of 89.9 kDa. The domain structure analysis revealed that the deduced protein has 2 Walker A domains, 2 Walker B domains, a Cdc48 domain, and a polyQ-binding domain. The mouse anti-HlVCP serum recognized a 97 kDa native protein in the salivary glands, midgut, and synganglion. RT–PCR analysis revealed that the native VCP was expressed throughout the developing stages and in tick organs. HlVCP silencing resulted in a decrease in tick body mass after blood feeding. This study not only contributes to a growing understanding of the ATPase gene family but also lays the groundwork for future studies on protein secretion and host–tick interaction. This study is the first report of the VCP gene from Chelicerata, which include spiders, scorpions, and ticks.

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A salivary cystatin, HlSC-1, from the ixodid tick Haemaphysalis longicornis play roles in the blood-feeding processes

Parasitology Research ◽

10.1007/s00436-009-1626-3 ◽

2009 ◽

Vol 106 (1) ◽

pp. 61-68 ◽

Cited By ~ 19

Author(s):

Kayoko Yamaji ◽

Naotoshi Tsuji ◽

Takeharu Miyoshi ◽

M. Khyrul Islam ◽

Takeshi Hatta ◽

...

Keyword(s):

Ixodid Tick ◽

Blood Feeding ◽

Haemaphysalis Longicornis

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Molecular characterization of a blood-induced serine carboxypeptidase from the ixodid tick Haemaphysalis longicornis

FEBS Journal ◽

10.1111/j.1742-4658.2007.05852.x ◽

2007 ◽

Vol 274 (13) ◽

pp. 3299-3312 ◽

Cited By ~ 27

Author(s):

Maki Motobu ◽

Naotoshi Tsuji ◽

Takeharu Miyoshi ◽

Xiaohong Huang ◽

M. K. Islam ◽

...

Keyword(s):

Molecular Characterization ◽

Ixodid Tick ◽

Haemaphysalis Longicornis ◽

Serine Carboxypeptidase

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Longistatin, a novel EF-hand protein from the ixodid tick Haemaphysalis longicornis, is required for acquisition of host blood-meals

International Journal for Parasitology ◽

10.1016/j.ijpara.2009.11.004 ◽

2010 ◽

Vol 40 (6) ◽

pp. 721-729 ◽

Cited By ~ 16

Author(s):

Anisuzzaman ◽

M. Khyrul Islam ◽

Takeharu Miyoshi ◽

M. Abdul Alim ◽

Takeshi Hatta ◽

...

Keyword(s):

Ixodid Tick ◽

Haemaphysalis Longicornis ◽

Ef Hand ◽

Host Blood ◽

Blood Meals

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A hemocyte-derived Kunitz–BPTI-type chymotrypsin inhibitor, HlChI, from the ixodid tick Haemaphysalis longicornis, plays regulatory functions in tick blood-feeding processes

Insect Biochemistry and Molecular Biology ◽

10.1016/j.ibmb.2012.09.005 ◽

2012 ◽

Vol 42 (12) ◽

pp. 925-934 ◽

Cited By ~ 7

Author(s):

M. Abdul Alim ◽

M. Khyrul Islam ◽

Anisuzzaman ◽

Takeharu Miyoshi ◽

Takeshi Hatta ◽

...

Keyword(s):

Ixodid Tick ◽

Blood Feeding ◽

Haemaphysalis Longicornis ◽

Chymotrypsin Inhibitor ◽

Regulatory Functions

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Molecular and reverse genetic characterization of serine proteinase-induced hemolysis in the midgut of the ixodid tick Haemaphysalis longicornis

Journal of Insect Physiology ◽

10.1016/j.jinsphys.2006.12.001 ◽

2007 ◽

Vol 53 (2) ◽

pp. 195-203 ◽

Cited By ~ 16

Author(s):

Takeharu Miyoshi ◽

Naotoshi Tsuji ◽

M. Khyrul Islam ◽

Xiaohong Huang ◽

Maki Motobu ◽

...

Keyword(s):

Genetic Characterization ◽

Serine Proteinase ◽

Ixodid Tick ◽

Haemaphysalis Longicornis ◽

Reverse Genetic

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Ultrafiltration Method for Plasma Protein Binding Studies and Its Limitations

Processes ◽

10.3390/pr9020382 ◽

2021 ◽

Vol 9 (2) ◽

pp. 382

Author(s):

Camelia-Maria Toma ◽

Silvia Imre ◽

Camil-Eugen Vari ◽

Daniela-Lucia Muntean ◽

Amelia Tero-Vescan

Keyword(s):

Protein Binding ◽

Plasma Protein Binding ◽

Plasma Protein ◽

Specific Binding ◽

Critical Role ◽

Volume Ratio ◽

Binding Studies ◽

Experimental Conditions ◽

Key Aspects

Plasma protein binding plays a critical role in drug therapy, being a key part in the characterization of any compound. Among other methods, this process is largely studied by ultrafiltration based on its advantages. However, the method also has some limitations that could negatively influence the experimental results. The aim of this study was to underline key aspects regarding the limitations of the ultrafiltration method, and the potential ways to overcome them. The main limitations are given by the non-specific binding of the substances, the effect of the volume ratio obtained, and the need of a rigorous control of the experimental conditions, especially pH and temperature. This review presents a variety of methods that can hypothetically reduce the limitations, and concludes that ultrafiltration remains a reliable method for the study of protein binding. However, the methodology of the study should be carefully chosen.

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