Activity and sequence characterization of two cysteine proteases in the digestive tract of the reduviid bug <i>Triatoma infestans</i>

Kollien, Astrid H.; Waniek, Peter J.; Nisbet, Alasdair J.; Billingsley, Peter F.; Schaub, Günter A.

doi:10.1111/j.0962-1075.2004.00504.x

Cited by 40 publications

(39 citation statements)

References 60 publications

(99 reference statements)

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“…The following degenerate primers were designed according to the amino acid residues flanking the His and Ser of the active site of serine proteinases (Sakanari et al 1989;Elvin et al 1993 (Kollien et al 2004b). Only the annealing parameters in a Hybaid (Omni Gene) thermal cycler (Hybaid, Ashford, Middx, UK) differed (54.2, 55.5, 56.9, 58.4, 59.3 and 59.8°C).…”

Section: Rna and Genomic Dna Isolationmentioning

confidence: 99%

“…To exclude Try1-cDNA, which contains an EcoRI restriction site at the positions 373-378 in the cDNA sequence, the PCR product was digested with EcoRI, separated on a 1% agarose-gel, and again a 470 bp fragment was cloned (Kollien et al 2004b). …”

Section: Rna and Genomic Dna Isolationmentioning

confidence: 99%

“…For the amplification of the 5¢-and 3¢-ends of the genes encoding trypsin and chymotrypsin, 5¢-and 3¢-RACE kits (Invitrogen, Karlsruhe, Germany) were used, following the manufacturer's protocol (Kollien et al 2004b).…”

Section: ¢-And 3¢-racementioning

confidence: 99%

“…For the subsequent amplification of target cDNA for Try2 the 5¢-GGAATTGCCCGTT GTGGGCC-3¢ forward primer and for Chy1 the 5¢-CTCGAGTACTATCCAGAAGCTCCTCT-3¢ forward primer were used with AUAP as the reverse primer. The resulting products, 516 bp for Try2 and 506 bp for Chy1, were cloned into the pGEM-T Easy and sequenced by MWG-BIOTECH (Kollien et al 2004b).…”

Section: ¢-And 3¢-racementioning

confidence: 99%

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Serine proteinases of the human body louse (Pediculus humanus): sequence characterization and expression patterns

et al. 2005

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After the previous characterization of one trypsin gene (Try1) of the human body louse Pediculus humanus, genes encoding a second trypsin (Try2) and a chymotrypsin (Chy1) have been cloned using degenerate serine proteinase primers and 5'- and 3'-RACE, and sequenced. The deduced 259 and 267 amino acid sequences of Try2 and Chy1 show an identity of 33%-40% to trypsinogens and chymotrypsinogens of other insects. Considering previously published partial sequences, P. humanus possesses at least one Try1 gene, five variants/isoforms of Try2 and six variants/isoforms of Chy1. The genomic DNA of Try2 contains three introns and Chy1 contains five introns. Using whole mount in situ hybridization, gene expression of Try1, Try2 and Chy1 has been localized not only in the distensible anterior region of the midgut of lice but sometimes also in the area following the distensible region. The Try2 gene was always expressed at much lower levels than Try1 or Chy1. This lower expression, the constitutive expression of Try1 and Chy1 at 1, 2, 6, 12 and 24 h after feeding of adults and the regional differences have been verified in quantitative real-time PCR.

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Section: Rna and Genomic Dna Isolationmentioning

confidence: 99%

Section: Rna and Genomic Dna Isolationmentioning

confidence: 99%

Section: ¢-And 3¢-racementioning

confidence: 99%

Section: ¢-And 3¢-racementioning

confidence: 99%

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Serine proteinases of the human body louse (Pediculus humanus): sequence characterization and expression patterns

et al. 2005

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“…In the beginning of the 1980s activities of cathepsin B and D, aminopeptidase and carboxypeptidase have been shown [16,17]. Later, when the appropriate molecular biology methods were available, different isoforms of genes from different triatomine species have been identified encoding cathepsin B, L and D and serine carboxypeptidase [18][19][20][21][22]. At the same time Borges et al [23] have demonstrated a significant increase of cathepsin D activity in Rhodnius prolixus infected with T. cruzi.…”

mentioning

confidence: 99%

Pathways of Insect Protein Digestion: Triatominae (Kissing Bugs)

Waniek¹

2014

Entomol Ornithol Herpetol

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EditorialThe majority of insects use serine proteinases like trypsin and chymotrypsin as main enzymes to digest their food [1]. These types of peptidases develop their maximum activity under alkaline to neutral pH conditions [2]. In some insect groups the intestinal lumen is acidic and the digestion switched to cysteine-like proteinases such as cathepsin D or L [3,4]. Some Coleoptera groups (e.g. Bruchidae, Tenebrionidae) developed a digestive system based on such proteinases [2,5]. But the major group using acid proteinases are the Hemiptera. This taxon uses cysteine proteinases because ancestral groups were using plant seeds containing serine proteinases inhibitors as a food source and therefore had to evolve their digestive system [6,7]. The conquest of this food source was a challenge for the digestive system [8] but it certainly contributed to the success of this insect group opening up new niches.One of the best analyzed hemipteran groups considering their digestion is the Triatominae (Heteroptera, Reduviidae) subfamily. Many species of this group -especially from the genera Triatoma, Rhodnius and Panstrongylus [9] -are medically important because they are able to transmit Trypanosoma cruzi (Kinetoplastida, Trypanosomatidae), the causative agent of Chagas disease [10]. In contrast to mosquitoes which develop from egg to adult in few weeks and frequently show low rates of parasitic infection, some long-lived triatomine species needs several months to complete their life cycle [1]. Besides this, kissing bugs takes huge amounts of blood from their vertebrate host at each nymph stage and, consequently usually show higher infection rates (up to about 80%) [1,[11][12][13][14][15]. In this insect group several digestive enzymes and their respective genes were identified and characterized. In the beginning of the 1980s activities of cathepsin B and D, aminopeptidase and carboxypeptidase have been shown [16,17]. Later, when the appropriate molecular biology methods were available, different isoforms of genes from different triatomine species have been identified encoding cathepsin B, L and D and serine carboxypeptidase [18][19][20][21][22]. At the same time Borges et al. [23] have demonstrated a significant increase of cathepsin D activity in Rhodnius prolixus infected with T. cruzi. However, despite the numerous studies concerning the Triatominae, many questions about their digestive system and its interactions with the parasite are still unanswered. For example, it is quite possible that there are further types of luminal digestive enzymes waiting to be discovered in triatomines. A recent transcriptome analysis of the R. prolixus intestine has revealed many genes related to digestive physiology including digestive enzymes lacking some amino acid residues important for enzymatic activity [24]. Furthermore, it would be very important to know whether or not microbiota plays a role in the digestion of triatomines and how strong the influence of T. cruzi on the digestive system might be. It would also be interesting ...

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Cathepsin B-like proteinase is involved in the decomposition of the adult fat body ofHelicoverpa armigera

Yang

Hou

Dong

et al. 2006

Arch. Insect Biochem. Physiol.

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Cathepsin B-like proteinase (HCB, EC 3.4.22.1) is expressed in Helicoverpa armigera oocytes and adult fat bodies. Previous work has revealed that HCB plays a key role in the degradation of yolk proteins during embryogenesis. This study investigated the function and regulatory activation of HCB in adult fat bodies during aging and oogenesis. The HCB transcript was detected at all stages from larval to adult fat bodies with Northern blot analysis. Pro-HCB was also detected in fat bodies at these stages with an immunoblot assay using a monoclonal antibody against HCB. However, mature HCB and its activity were only detected in fat bodies of pre-adults and adults. This evidence suggested that HCB is regulated post-translationally by activation of the pro-enzyme during the pupa-adult metamorphosis. The activation of HCB was coupled with the expression of hormone receptor 3 (HHR3), and was up-regulated by the ecdysteroid agonist, RH-2485, suggesting that HCB activation is related to the ecdysone regulatory system. The decomposition of the adult fat bodies during aging and oogenesis was found to occur via programmed cell death, in which HCB took part.

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Activity and sequence characterization of two cysteine proteases in the digestive tract of the reduviid bug Triatoma infestans

Cited by 40 publications

References 60 publications

Serine proteinases of the human body louse (Pediculus humanus): sequence characterization and expression patterns

Serine proteinases of the human body louse (Pediculus humanus): sequence characterization and expression patterns

Pathways of Insect Protein Digestion: Triatominae (Kissing Bugs)

Cathepsin B-like proteinase is involved in the decomposition of the adult fat body ofHelicoverpa armigera

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