Molecular cloning, mRNA expression and enzymatic characterization of cathepsin F from olive flounder (Paralichthys olivaceus)

Ahn, Sang Jung; Kim, Na Young; Seo, Jung Soo; Je, Ju Eun; Sung, Ji Hea; Lee, Sang-Hwan; Kim, Moo‐Sang; Kim, Joong Kyun; Chung, Joon Ki; Lee, Hyung Ho

doi:10.1016/j.cbpb.2009.06.005

Cited by 21 publications

(14 citation statements)

References 50 publications

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“…To determine the inhibitory activity of PoCystatin B against the piscine cathepsins and bovine cathepsin B (C6286, Sigma-Aldrich, USA), the residual enzyme activity following incubation with PoCystatin B was measured using Z-Phe-Arg-AMC as the substrate for cathepsins L (pH 7.5, Ahn et al, 2010), F (pH 7.5, Ahn et al, 2009), and X (pH 5.0, Ahn et al, 2008), Z-Val-Val-Arg-AMC as the substrate for cathepsin S (pH 8.0, Kim et al, 2010), Z-Gly-Pro-Arg-AMC as the substrate for cathepsin K (pH 8.0, Je et al, 2009), and Z-Arg-Arg-AMC as the substrate for cathepsin B (pH 5.0) in 100 mM sodium acetate (pH 4-5.5) or 100 mM Tris (pH 7.5-8.5) buffers. The amount of 7-amido-4-methylcoumarin (AMC) released after enzymatic cleavage was measured using a Microplate Fluorometer (Packard Co. USA) at an excitation wavelength of 380 nm and emission wavelength of 460 nm.…”

Section: Inhibitor Assaymentioning

confidence: 99%

Olive flounder (Paralichthys olivaceus) cystatin B: Cloning, tissue distribution, expression and inhibitory profile of piscine cystatin B

Ahn¹,

Bak²,

Park³

et al. 2013

Comparative Biochemistry and Physiology Part B: Biochemistry an

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Section: Inhibitor Assaymentioning

confidence: 99%

Olive flounder (Paralichthys olivaceus) cystatin B: Cloning, tissue distribution, expression and inhibitory profile of piscine cystatin B

Ahn¹,

Bak²,

Park³

et al. 2013

Comparative Biochemistry and Physiology Part B: Biochemistry an

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“…Cathepsin F (GenBank accession no. ACC86111) was recently published from olive flounder (Ahn et al, 2009). The published cathepsin F gene was identical to the isolated one except for two-amino acid changes in the mature region (Y455N and H457Y).…”

Section: Resultsmentioning

confidence: 91%

“…The cathepsin F protein showed 99% amino acid sequence identity with published flounder cathepsin F (Ahn et al, 2009). When compared to other cathepsin F, the full-length amino acid identities were 74% with Atlantic salmon (NP_001133678), 73% with zebrafish (NP_001071036), 53% with cattle (NP_001068884), 52% with dog (XP_533219) and mouse (NP_063914) and 49% with human (NP_003784) (Fig.…”

Section: Alignments and Phylogenetic Tree Constructionmentioning

confidence: 90%

“…The high expression level of the liver tissue which is related to the immune response in fish, suggests that cathepsin F may have the immune-related functions. Ahn et al (2009) reported that the flounder cathepsin F plays an important role in the epidermal mucus and fish innate immune system. To determine the stage of development in which transcription of cathepsin F occurs, we analyzed at the five differ- Semi-quantitative analysis was performed on total RNA isolated from the internal organs of adult olive flounder.…”

Section: Alignments and Phylogenetic Tree Constructionmentioning

confidence: 99%

“…In recent years, a number of cathepsin F have been discovered and described at the molecular level from a number of sources (Wang et al, 1998;Nägler et al, 1999;Deussing et al, 2000;Joo et al, 2007). In fish, cathepsin F is also known to be involved in several important functions such as oocyte maturation (Fabra and Cerdá, 2004), development of the zebrafish retina (Vihtelic et al, 2005), and bactericidal role in epidermal mucus (Ahn et al, 2009). In the present study we aimed to characterize the cathepsin F cDNA isolated from the olive flounder Paralichthys olivaceus, and to investigate the expression of cathepsin F transcripts in various tissues of adult as well as in whole-body larvae at early developmental stages.…”

Section: Introductionmentioning

confidence: 98%

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Molecular cloning and characterization of cathepsin F gene from olive flounder Paralichthys Olivaceus

et al. 2010

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We isolated a homologue of cathepsin F from cDNA library of olive flounder liver. A 2,077 kb full-length cDNA encoding a predicted polypeptide of 474 amino acids was sequenced. The flounder cathepsin F exhibits a domain structure typical for papain-like cysteine proteases, a 17 amino acid N-terminal hydrophobic signal sequence followed by an extraordinarily long propeptide of 244 amino acids and the domain of the mature protease comprising 213 amino acids. The mature region contains all features characteristic of a papain-like cysteine protease, including the highly conserved cysteine, histidine and asparagine residues of the 'catalytic triad'. The cathepsin F protein showed 49−99% amino acid sequence identity with other known cathepsin F sequences. An in vivo expression study showed that cathepsin F mRNA was expressed predominantly in brain, liver, eye and heart, and moderately in other tissues. The accumulation of cathepsin F mRNA in early stage of development increased with development. This expression pattern suggests that flounder cathepsin F has been implicated in the growth and reproduction regulation.

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Olive Flounder (Paralichthys olivaceus) Cystatin C: Cloning, mRNA Expression, and Enzymatic Characterization of Olive Flounder Cystatin C

Ahn

Bak

Park

et al. 2013

Appl Biochem Biotechnol

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Cystatins are endogenous inhibitors of mammalian lysosomal cysteine proteinases, such as cathepsins B, L, H, and S. Cystatin C belongs to the type 2 cystatin family. In this study, the 751-bp cystatin C cDNA (PoCystatin C) of olive flounder (Paralichthys olivaceus) was cloned by screening from the olive flounder cDNA library. The mRNA expression of the PoCystatin C gene was examined in various tissues from normal and lipopolysaccharide (LPS)-stimulated olive flounder by RT-PCR and was compared with inflammatory cytokines IL-1β, IL-6, and IL-8. PoCystatin C transcripts ubiquitously existed in all normal and LPS-stimulated tissues that were tested. The recombinant PoCystatin C protein was expressed in Escherichia coli BL21(DE3) in pCold™ TF DNA expression vector as a 70-kDa fusion protein. The protease inhibitory activities of recombinant PoCystatin C toward papain cysteine protease, piscine cathepsins (L, S, K, F, and X), and bovine cathepsin B were measured with the synthetic fluorogenic peptide substrates. PoCystatin C tightly inhibited papain cysteine protease, whereas cathepsins L, S, K, F, X, and B were inhibited with lower affinities. Our results indicate that the P. olivaceus cystatin C is a homolog of mammalian cystatin C due to its sequence, structure, tissue expression, and biochemical activity.

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Molecular cloning, mRNA expression and enzymatic characterization of cathepsin F from olive flounder (Paralichthys olivaceus)

Cited by 21 publications

References 50 publications

Olive flounder (Paralichthys olivaceus) cystatin B: Cloning, tissue distribution, expression and inhibitory profile of piscine cystatin B

Olive flounder (Paralichthys olivaceus) cystatin B: Cloning, tissue distribution, expression and inhibitory profile of piscine cystatin B

Molecular cloning and characterization of cathepsin F gene from olive flounder Paralichthys Olivaceus

Olive Flounder (Paralichthys olivaceus) Cystatin C: Cloning, mRNA Expression, and Enzymatic Characterization of Olive Flounder Cystatin C

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