Cloning and molecular characterization of a cubilin-related serine proteinase from the hard tick Haemaphysalis longicornis

Miyoshi, Takeharu; Tsuji, Naotoshi; Islam, M. Khyrul; Kamio, Tsugihiko; Fujisaki, Kozo

doi:10.1016/j.ibmb.2004.04.004

Cited by 41 publications

(42 citation statements)

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“…But in the blood-feeding ticks, only aspartic and cysteine proteinases have been characterized enzymatically [3,13,14,16]. In the previous study, we have identified HlSP gene from the hard tick H. longicornis as a member of the serine proteinase gene family [9]. The deduced amino acid sequence of HlSP showed the highest similarity with the midgut serine proteinase of Rhipicephalus appendiculatus (AAC79567) and also showed similarity with mammalian pancreatic elastases.…”

Section: Discussionmentioning

confidence: 98%

“…Enzyme assay: Enzyme assay was performed as previously described [9]. Briefly, substrate specificity assays with the rHlSP were performed in a standard reaction mixture containing 25 mM citric acid/50 mM sodium phosphate pH 5.0, 5 mM dithiothreitol and 200 µM of chromogenic or fluorogenic substrates.…”

Section: Methodsmentioning

confidence: 99%

“…In particular, H. longicornis serves as a vector of Theileria orientalis, which is the most economically important protozoan infectious disease of cattle in Asia [6]. In contrast to the extensive reports on proteinase activities in haematophagous insects, very little is known about these activities in the hard tick H. longicornis despite of its high vector potentiality.Recently, we cloned and partially characterized a gene that encodes a functional serine proteinase from the hard tick H. longicornis [9]. The H. longicornis serine proteinase (HlSP, Genbank accession number: AB127388) with a signal sequence was composed of complement C1r/C1s, Uegf, and bone morphogenic protein-1 (CUB) domain, low-density lipoprotein receptor class A (LDL) domain and catalytic domain and has a molecular mass of 50 kDa and a pI of 8.2.…”

mentioning

confidence: 99%

“…Sci. 66(10): 1195-1198, 2004 Ticks and tick-borne diseases are of extreme importance throughout the world and the associated economic losses are immense. Reliance on acaricides for tick control has led to serious problems which include development of resistance to acaricides that created control problems [5], and increased the threat to the wide-spread occurrence of the pathogens associated with ticks.…”

mentioning

confidence: 99%

“…Recently, we cloned and partially characterized a gene that encodes a functional serine proteinase from the hard tick H. longicornis [9]. The H. longicornis serine proteinase (HlSP, Genbank accession number: AB127388) with a signal sequence was composed of complement C1r/C1s, Uegf, and bone morphogenic protein-1 (CUB) domain, low-density lipoprotein receptor class A (LDL) domain and catalytic domain and has a molecular mass of 50 kDa and a pI of 8.2.…”

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Enzymatic Characterization of a Cubilin-Related Serine Proteinase from the Hard Tick Haemaphysalis longicornis

et al. 2004

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ABSTRACT. In the present study, we performed enzymatic characterization of Haemaphysalis longicornis serine proteinase (HlSP) with a view to shed light on the mechanisms of blood digestion in the hard ticks. Escherichia coli-expressed recombinant HlSP (rHlSP) was shown to potently hydrolyze the synthetic substrates Bz-(DL)-Arg-pNA, Z-Ala-Ala-Leu-pNA and Suc-Ala-Ala-Ala-pNA and yielded an activity of 31.5, 88.2 and 18.3 µmol/min/mg protein, respectively at an optimum temperature of 25°C. However, the enzyme showed little activity to hydrolyze the substratese Suc-Arg-Pro-Phe-His-Leu-Leu-Val-Tyr-MCA and Pyr-Phe-Leu-pNA. The optimum pH for the enzyme was shown to be 4.0 to 5.0. Several inhibitors such as antipain, leupeptin and phenylmethylsulfonyl fluoride (PMSF), specific for serine proteinase were shown to inhibit enzyme activity by 20-82%, while E-64 (specific for cysteine proteinases) and pepstatinA (specific for aspartic proteinases) had shown only little inhibitory effects on it. This is the first report on enzymatic characterization of a functional serine proteinase from the hard ticks. KEY WORDS: enzyme activity, Haemaphysalis longicornis, serine proteinase.J. Vet. Med. Sci. 66(10): 1195-1198, 2004 Ticks and tick-borne diseases are of extreme importance throughout the world and the associated economic losses are immense. Reliance on acaricides for tick control has led to serious problems which include development of resistance to acaricides that created control problems [5], and increased the threat to the wide-spread occurrence of the pathogens associated with ticks. In addition, chemical residues in animal products, and the effects of such chemicals on the environment, must be considered when new acaricides are developed.Proteolytic enzymes may represent interesting target molecules for drug design because of its involvement in the mediation of a wide range of cellular processes such as protein metabolism and the processing of precursor proteins in the living organisms [11]. In haematophagous ticks, blood digestion in the midgut is thought to be the most essential process because it is the source of energy as well as the most critical interface between the ticks and the pathogens that transmitted by ticks. Studies have reported that aspartic and cysteine proteinases exert proteolytic role, in the midguts [8,13] of the hard tick Boophilus microplus.The hard tick Haemaphysalis longicornis is distributed in a vast area of Eurasia, including Japan [6], where it transmits a wide range of pathogens, including viruses, rickettsia and protozoan parasites, causing important human and animal diseases [17]. In particular, H. longicornis serves as a vector of Theileria orientalis, which is the most economically important protozoan infectious disease of cattle in Asia [6]. In contrast to the extensive reports on proteinase activities in haematophagous insects, very little is known about these activities in the hard tick H. longicornis despite of its high vector potentiality.Recently, we cloned and partially characteri...

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Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Enzymatic Characterization of a Cubilin-Related Serine Proteinase from the Hard Tick Haemaphysalis longicornis

et al. 2004

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Molecular Targets to Impair Blood Meal Processing in Ticks

Kopáček¹,

Perner²,

Sojka³

et al. 2018

Ectoparasites

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RNA interference technology in crop protection against arthropod pests, pathogens and nematodes

Zotti

Santos

Cagliari

et al. 2018

Pest Management Science

306

243

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Scientists have made significant progress in understanding and unraveling several aspects of double-stranded RNA (dsRNA)-mediated gene silencing during the last two decades. Now that the RNA interference (RNAi) mechanism is well understood, it is time to consider how to apply the acquired knowledge to agriculture and crop protection. Some RNAi-based products are already available for farmers and more are expected to reach the market soon. Tailor-made dsRNA as an active ingredient for biopesticide formulations is considered a raw material that can be used for diverse purposes, from pest control and bee protection against viruses to pesticide resistance management. The RNAi mechanism works at the messenger RNA (mRNA) level, exploiting a sequence-dependent mode of action, which makes it unique in potency and selectivity compared with conventional agrochemicals. Furthermore, the use of RNAi in crop protection can be achieved by employing plant-incorporated protectants through plant transformation, but also by non-transformative strategies such as the use of formulations of sprayable RNAs as direct control agents, resistance factor repressors or developmental disruptors. In this review, RNAi is presented in an agricultural context (discussing products that have been launched on the market or will soon be available), and we go beyond the classical presentation of successful examples of RNAi in pest-insect control and comprehensively explore its potential for the control of plant pathogens, nematodes and mites, and to fight against diseases and parasites in beneficial insects. Moreover, we also discuss its use as a repressor for the management of pesticide-resistant weeds and insects. Finally, this review reports on the advances in non-transformative dsRNA delivery and the production costs of dsRNA, and discusses environmental considerations.

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Cloning and molecular characterization of a cubilin-related serine proteinase from the hard tick Haemaphysalis longicornis

Cited by 41 publications

References 30 publications

Enzymatic Characterization of a Cubilin-Related Serine Proteinase from the Hard Tick Haemaphysalis longicornis

Enzymatic Characterization of a Cubilin-Related Serine Proteinase from the Hard Tick Haemaphysalis longicornis

Molecular Targets to Impair Blood Meal Processing in Ticks

RNA interference technology in crop protection against arthropod pests, pathogens and nematodes

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