Characterization and Expression of Trypsinogen and Trypsin in Medaka Testis

Rajapakse, Senaka; Ogiwara, Katsueki; Takahashi, Takayki

doi:10.2108/zs140111

Cited by 8 publications

(3 citation statements)

References 44 publications

(51 reference statements)

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“…Similarly, in medaka, the trypsinogen gene is expressed primarily in the mesentery, including the pancreas, and weakly in the intestines and testes. These results agree with a previous report 13 . It can be concluded that gene expression of the three pactacins corresponds closely to that of trypsinogen genes in seahorse and medaka.…”

Section: Resultssupporting

confidence: 94%

Pactacin is a novel digestive enzyme in teleosts

Kawaguchi

Okazawa

Imafuku

et al. 2021

Sci Rep

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Generally, animals extract nutrients from food by degradation using digestive enzymes. Trypsin and chymotrypsin, one of the major digestive enzymes in vertebrates, are pancreatic proenzymes secreted into the intestines. In this investigation, we report the identification of a digestive teleost enzyme, a pancreatic astacin that we termed pactacin. Pactacin, which belongs to the astacin metalloprotease family, emerged during the evolution of teleosts through gene duplication of astacin family enzymes containing six cysteine residues (C6astacin, or C6AST). In this study, we first cloned C6AST genes from pot-bellied seahorse (Hippocampus abdominalis) and analyzed their phylogenetic relationships using over 100 C6AST genes. Nearly all these genes belong to one of three clades: pactacin, nephrosin, and patristacin. Genes of the pactacin clade were further divided into three subclades. To compare the localization and functions of the three pactacin subclades, we studied pactacin enzymes in pot-bellied seahorse and medaka (Oryzias latipes). In situ hybridization revealed that genes of all three subclades were commonly expressed in the pancreas. Western blot analysis indicated storage of pactacin pro-enzyme form in the pancreas, and conversion to the active forms in the intestine. Finally, we partially purified the pactacin from digestive fluid, and found that pactacin is novel digestive enzyme that is specific in teleosts.

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Section: Resultssupporting

confidence: 94%

Pactacin is a novel digestive enzyme in teleosts

Kawaguchi

Okazawa

Imafuku

et al. 2021

Sci Rep

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“…Another recombinant fish trypsin partially characterized is the one obtained from Medaka (Oryzias latipes) and produced in E. coli. The Km and catalytic efficiency of this recombinant enzyme were 0.10 mM and 7.15 s −1 mM −1 , respectively, using Boc-Gln-Ala Arg-MCA as substrate (Rajapakse et al 2014). It is worth mentioning that a complete biochemical and kinetic characterization has not been performed on most of the recombinant fish trypsins, making their comparison with native trypsins difficult.…”

Section: Features Of Recombinant Fish Trypsinsmentioning

confidence: 99%

Fish trypsins: potential applications in biomedicine and prospects for production

Cruz¹,

Álvarez‐González²,

Peña³

et al. 2018

3 Biotech

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In fishes, trypsins are adapted to different environmental conditions, and the biochemical and kinetic properties of a broad variety of native isoforms have been studied. Proteolytic enzymes remain in high demand in the detergent, food, and feed industries; however, our analysis of the literature showed that, in the last decade, some fish trypsins have been studied for the synthesis of industrial peptides and for specific biomedical uses as antipathogenic agents against viruses and bacteria, which have been recently patented. In addition, innovative strategies of trypsin administration have been studied to ensure that trypsins retain their properties until they exert their action. Biomedical uses require the production of high-quality enzymes. In this context, the production of recombinant trypsins is an alternative. For this purpose, -based systems have been tested for the production of fish trypsins; however,-based systems also seem to show great potential in the production of fish trypsins with higher production quality. On the other hand, there is a lack of information regarding the specific structures, biochemical and kinetic properties, and characteristics of trypsins produced using heterologous systems. This review describes the potential uses of fish trypsins in biomedicine and the enzymatic and structural properties of native and recombinant fish trypsins obtained to date, outlining some prospects for their study.

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“…The POP family is a new group of serine peptidases and different from the classic serine proteases, trypsin (Rajapakse, Ogiwara, & Takahashi, 2014) and subtilisin (Fernández, Daleo, & Guevara, 2015), in several structural features and catalytic behaviors (Polgar, 1994(Polgar, , 2002aRennex, Hemmings, Hofsteenge, & Stone, 1991;Szeltner et al, 2003). They regulate the activity of biologically active peptides and peptide hormones, and they are implicated in diseases, including amnesia, depression, diabetes, and trypanosomiasis (Rea & Fülöp, 2006).…”

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confidence: 99%

Exploration of the chlorpyrifos escape pathway from acylpeptide hydrolases using steered molecular dynamics simulations

Wang¹,

Jin²,

Wang³

et al. 2015

Journal of Biomolecular Structure and Dynamics

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Acylpeptide hydrolases (APH) catalyze the removal of an N-acylated amino acid from blocked peptides. APH is significantly more sensitive than acetylcholinesterase, a target of Alzheimer's disease, to inhibition by organophosphorus (OP) compounds. Thus, OP compounds can be used as a tool to probe the physiological functions of APH. Here, we report the results of a computational study of molecular dynamics simulations of APH bound to the OP compounds and an exploration of the chlorpyrifos escape pathway using steered molecular dynamics (SMD) simulations. In addition, we apply SMD simulations to identify potential escape routes of chlorpyrifos from hydrolase hydrophobic cavities in the APH-inhibitor complex. Two previously proposed APH pathways were reliably identified by CAVER 3.0, with the estimated relative importance of P1 > P2 for its size. We identify the major pathway, P2, using SMD simulations, and Arg526, Glu88, Gly86, and Asn65 are identified as important residues for the ligand leaving via P2. These results may help in the design of APH-targeting drugs with improved efficacy, as well as in understanding APH selectivity of the inhibitor binding in the prolyl oligopeptidase family.

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Characterization and Expression of Trypsinogen and Trypsin in Medaka Testis

Cited by 8 publications

References 44 publications

Pactacin is a novel digestive enzyme in teleosts

Pactacin is a novel digestive enzyme in teleosts

Fish trypsins: potential applications in biomedicine and prospects for production

Exploration of the chlorpyrifos escape pathway from acylpeptide hydrolases using steered molecular dynamics simulations

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