Purification and Characterization of Chymotrypsin From Penaeus Vannamei (Crustacea: Decapoda)

Hernández-Cortés, Patricia; Whitaker, John R.; García‐Carreño, Fernando

doi:10.1111/j.1745-4514.1997.tb00202.x

Cited by 54 publications

(33 citation statements)

References 25 publications

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“…In the mammalian eye, serine proteases play multiple roles in development processes (Masos et al, 2000), such as acting as an avoidance cue in axonal guidance during development (Baird and Rapier, 1995). In crustaceans, serine proteases and their inhibitors have been shown to function in digestion (Vanwormhoudt et al, 1995;Klein et al, 1996;Roy et al, 1996;Hernandez-Cortes et al, 1997) and in immunity (Soderhall and Cerenius, 1998;Kanost, 1999). However, the role of serine proteases in crustacean nervous systems is virtually unexplored.…”

Section: Introductionmentioning

confidence: 97%

A CUB‐serine protease in the olfactory organ of the spiny lobster Panulirus argus

et al. 2001

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csp, a gene encoding a protein with high sequence identity to trypsinlike serine protease and CUB domains, was identified from a cDNA library from the olfactory organ (antennular lateral flagellum) of the spiny lobster Panulirus argus. The full-length cDNA sequence of csp is 1801 bp, encoding a protein of 50.25 kD, with three domains: signal peptide, trypsinlike serine protease, and CUB (named for a class of compounds including Complement subcomponents Clr/Cls, Uegf, and Bone morphogenic protein-1). RT-PCR, Northern blots, and immunoblots showed that csp is predominantly expressed in the lateral flagellum and eyestalk. Immunocytochemistry showed that Csp is present in olfactory (aesthetasc) sensilla around auxiliary cells (glia that surround the inner dendrites of olfactory receptor neurons, ORNs) and ORN outer dendrites. We propose that Csp is expressed and secreted by auxiliary cells, associates with ORN cell membranes or extracellular matrix via the CUB domain, and has trypsinlike activity. In the eyestalk, Csp is associated with cells surrounding axons between neuropils of the eyestalk ganglia. Possible functions in the olfactory organ and eyestalk are discussed. To our knowledge, this is the first report from any olfactory system of a gene encoding a protein with serine protease and CUB domains.

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Section: Introductionmentioning

confidence: 97%

A CUB‐serine protease in the olfactory organ of the spiny lobster Panulirus argus

et al. 2001

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“…It is synthesised in large quantities and may represent up to one-third of the soluble proteins in the digestive gland (Ceccaldi, 1998). Many investigations confirmed high trypsin activities in different crustacean species (e.g., Murhty and Saxena, 1979;Garcia-Carreño et al, 1994Johnston et al, 1995;Hernández-Cortés et al, 1997;Le Moullac et al, 1997). However, preliminary but repeated enzyme analysis of the midgut glands of C. crangon as well as C. allmani showed unusually low or even no trypsin activities, while in the same samples, total proteolytic activities were high (Saborowski, unpublished).…”

Section: Introductionmentioning

confidence: 99%

Cysteine proteinases substitute for serine proteinases in the midgut glands of Crangon crangon and Crangon allmani (Decapoda: Caridea)

Teschke

Saborowski

2005

Journal of Experimental Marine Biology and Ecology

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The utilization of dietary proteins in crustaceans is facilitated by a set of peptide hydrolases which are often dominated by btrypsin-likeQ serine proteinases. As expected, the North Sea shrimps Crangon crangon and Crangon allmani showed in their midgut glands high proteolytic activities. However, the majority of animals lacked trypsin and chymotrypsin. Conversely, a minority of about 10% of the animals had elevated trypsin activities. The appearance of trypsin was neither related to the mode of feeding nor to the nutritive state of the animals. When present, trypsin was expressed in both species as a single isoform of apparently 20 kDa. The lack of serine proteinases was also confirmed by inhibitor assays. AEBSF, a serine proteinase inhibitor, slightly reduced total proteinase activity by less than 10%. In contrast E 64, a cysteine proteinase inhibitor, caused a reduction of more than 70% of total proteinase activity, indicating that a substantial share of proteolytic activity is caused by cysteine proteinases. Cathepsin L-like proteinases were identified as major cysteine proteinases.A comparison with the eucarid crustaceans Pandalus montagui, Pagurus bernhardus, Cancer pagurus and Euphausia superba showed a similar high level of total proteinase activity in all species. Trypsin, however, varied significantly between species showing lowest activities in Caridea and the highest activity in E. superba. E 64 suppressed total proteinase activity by more than 70% in Crangon species but not in C. pagurus and E. superba. In contrast, the serine proteinase inhibitor AEBSF had only little effect in Caridea but was most effective in P. bernhardus, C. pagurus and E. superba. The results may indicate different traits of food utilization strategies in some eucarid crustaceans. Caridea may express predominantly cysteine proteinase, while in Anomura, Brachyura and Euphausiacea, serine proteinases may prevail. D

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“…In the digestive tract of decapods, serine proteinases, such as trypsin and chymotrypsin (Galgani et al 1985;Hernández-Cortés et al 1999;Hernández-Cortés et al 1997); cysteine proteinases, such as cathepsin L and cathepsin B (Aoki et al 2004;Le Boulay et al 1998;Teschke and Saborowski 2005); and metallo-proteinses, such as astacin (Titani et al 1987;Zwilling and Stöcker 1997) were studied most intensively, while aspartic proteinases remained unnoticed. In clawed lobsters (Homaridae), digestive proteolytic enzymes have been reported since the early 1970s (Brockerhoff et al 1970;Hoyle 1973), and, unlike other decapods, these proteinases show highest activity at acid pH (Baker and Gibson 1977;Glass and Stark 1994).…”

Section: Introductionmentioning

confidence: 99%

Aspartic Cathepsin D Endopeptidase Contributes to Extracellular Digestion in Clawed Lobsters Homarus americanus and Homarus gammarus

et al. 2010

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Acid digestive proteinases were studied in the gastric fluids of two species of clawed lobster (Homarus americanus and Homarus gammarus). An active protein was identified in both species as aspartic proteinase by specific inhibition with pepstatin A. It was confirmed as cathepsin D by mass mapping, N-terminal, and full-length cDNA sequencing. Both lobster species transcribed two cathepsin D mRNAs: cathepsin D1 and cathepsin D2. Cathepsin D1 mRNA was detected only in the midgut gland, suggesting its function as a digestive enzyme. Cathepsin D2 mRNA was found in the midgut gland, gonads, and muscle. The deduced amino acid sequence of cathepsin D1 and cathepsin D2 possesses two catalytic DTG active-site motifs, the hallmark of aspartic proteinases. The putatively active cathepsin D1 has a molecular mass of 36.4 kDa and a calculated pI of 4.14 and possesses three potential glycosylation sites. The sequences showed highest similarities with cathepsin D from insects but also with another crustacean cathepsin D. Cathepsin D1 transcripts were quantified during a starvation period using real-time qPCR. In H. americanus, 15 days of starvation did not cause significant changes, but subsequent feeding caused a 2.5-fold increase. In H. gammarus, starvation caused a 40% reduction in cathepsin D1 mRNA, and no effect was observed with subsequent feeding.

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Purification and Characterization of Chymotrypsin From Penaeus Vannamei (Crustacea: Decapoda)

Cited by 54 publications

References 25 publications

A CUB‐serine protease in the olfactory organ of the spiny lobster Panulirus argus

A CUB‐serine protease in the olfactory organ of the spiny lobster Panulirus argus

Cysteine proteinases substitute for serine proteinases in the midgut glands of Crangon crangon and Crangon allmani (Decapoda: Caridea)

Aspartic Cathepsin D Endopeptidase Contributes to Extracellular Digestion in Clawed Lobsters Homarus americanus and Homarus gammarus

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