Three Kazal-type serine proteinase inhibitors from the red swamp crayfish Procambarus clarkii and the characterization, function analysis of hcPcSPI2

Li, Xin-Cang; Zhang, Ranran; Sun, Rong-Rong; Lan, Jiang-Feng; Zhao, Xiao‐Fan; Wang, Jin-Xing

doi:10.1016/j.fsi.2010.02.011

Cited by 27 publications

(13 citation statements)

References 35 publications

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“…The recombinant Sp TubeDD protein with a glutathione S-transferase (GST) tag and a GST protein was purified using glutathione Sepharose 4B chromatography (GenScript, USA) according to the manufacturer’s instructions. Both Sp MyD88DD and Sp PelleDD proteins with an N-terminal His tag were harvested by His Bind resin chromatography (Novagen, USA) according to a previous method [30].…”

Section: Methodsmentioning

confidence: 99%

Identification, Characterization, and Functional Analysis of Tube and Pelle Homologs in the Mud Crab Scylla paramamosain

Zhang

Zhou

et al. 2013

PLoS ONE

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Tube and Pelle are essential components in Drosophila Toll signaling pathway. In this study, we characterized a pair of crustacean homologs of Tube and Pelle in Scylla paramamosain, namely, SpTube and SpPelle, and analyzed their immune functions. The full-length cDNA of SpTube had 2052 bp with a 1578 bp open reading frame (ORF) encoding a protein with 525 aa. A death domain (DD) and a kinase domain were predicted in the deduced protein. The full-length cDNA of SpPelle had 3825 bp with a 3420 bp ORF encoding a protein with 1140 aa. The protein contained a DD and a kinase domain. Two conserved repeat motifs previously called Tube repeat motifs present only in insect Tube or Tube-like sequences were found between these two domains. Alignments and structure predictions demonstrated that SpTubeDD and SpPelleDD significantly differed in sequence and 3D structure. Similar to TubeDD, SpTubeDD contained three common conserved residues (R, K, and R) on one surface that may mediate SpMyD88 binding and two common residues (A and A) on the other surface that may contribute to Pelle binding. By contrast, SpPelleDD lacked similar conservative residues. SpTube, insect Tube-like kinases, and human IRAK4 were found to be RD kinases with an RD dipeptide in the kinase domain. SpPelle, Pelle, insect Pelle-like kinases, and human IRAK1 were found to be non-RD kinases lacking an RD dipeptide. Both SpTube and SpPelle were highly expressed in hemocytes, gills, and hepatopancreas. Upon challenge, SpTube and SpPele were significantly increased in hemocytes by Gram-negative or Gram-positive bacteria, whereas only SpPelle was elevated by White Spot Syndrome Virus. The pull-down assay showed that SpTube can bind to both SpMyD88 and SpPelle. These results suggest that SpTube, SpPelle, and SpMyD88 may form a trimeric complex involved in the immunity of mud crabs against both Gram-negative and Gram-positive bacteria.

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Section: Methodsmentioning

confidence: 99%

Identification, Characterization, and Functional Analysis of Tube and Pelle Homologs in the Mud Crab Scylla paramamosain

Zhang

Zhou

et al. 2013

PLoS ONE

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“…Then, the cDNAs were synthesized as the templates with the total RNA using the PrimeScriptRT reagent kit (Takara) according the manufacturer's instructions. To detect the expression profiles of CYP1A and 3A at mRNA level, quantitative real-time polymerase chain reaction (qRT-PCR) was performed following the methods described in a previous report (Li et al, 2010). The primers, F1 (5¢-CAT CCC TTT CTT GCG TAT CCT-3¢) and R1 (5¢-CGT TTG AGT TCT CGT CCA GTT T-3¢) for CYP1A (Accession no.…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

Effects of enrofloxacin on cytochromes P4501A and P4503A in Carassius auratus gibelio (crucian carp)

Li²,

Sun³

et al. 2011

Vet Pharm & Therapeutics

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Currently, although enrofloxacin (EF) as a widely used veterinary medicine has begun to apply to treating fish bacterial infections, the researches on the effects of EF on their main drug metabolic enzymes are limited. To investigate the effects of EF on fish cytochromes P450 (CYPs) 1A and 3A, the enzymatic activities and expressions (mRNA and protein) of crucian carp CYP1A and CYP3A after EF administration were examined. For CYP1A, in the in vivo experiments, EF exhibited potent inhibition on the CYP1A-related ethoxyresorufin-O-deethylase (EROD) activity, as well as CYP1A expressions at both protein and mRNA levels, at 24 h after administration with different EF dosages (3, 10, 30, and 60 mg/kg); Furthermore, CYP1A enzymatic activity and expressions at both protein and mRNA levels decreased more with increasing EF dosages. Additionally, the in vitro experimental results showed that, after incubated with microsomes, EF did not change the EROD activity through interacting directly with CYP1A. For CYP3A, the in vitro and in vivo experimental results demonstrated that EF could inhibit the CYP3A-related erythromycin N-demethylase activity in a time- and dose-dependent manner, while it did not suppress CYP3A expressions at both protein and mRNA levels after administration with EF for a short period (no more than 24 h); however, after injection with EF at a high dose (10 mg/kg) for a long period, the CYP3A protein and mRNA reached their lowest levels at 96 and 48 h, respectively. These results indicate that EF can suppress CYP1A expressions in a dose-dependent manner, thereby inhibiting further its catalytic activity; meanwhile, both the interactions of EF with CYP3A and the expressions decrease (protein and mRNA) caused by EF contribute to the CYP3A inhibition.

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“…In the Chinese white shrimp F. chinensis, four Kazal inhibitors (FcSPI-1, FcSPI-2, FcSPI-3 and FcSPI-4) were identified in the hepatopancreas [22]. Similar polymorphisms were also found in the freshwater crayfish P. leniusculus [45] and the red swamp crayfish P. clarkii [23,24]. Therefore, we cannot exclude the possibility that other KPI proteins may exist in M. nipponense and E. sinensis in addition to Man-KPI and Ers-KPI.…”

Section: Discussionmentioning

confidence: 57%

“…In crustaceans, numerous KPIs have been identified in recent years, most notably in the species Penaeus monodon [19,20], Litopenaeus vannamei [21], Fenneropenaeus chinensis [11,22] and Procambarus clarkii [23,24]. These KPIs are all expressed outside the male reproductive system and are believed to be involved in the innate immune response [25].…”

Section: Introductionmentioning

confidence: 99%

Two Kazal-type protease inhibitors from Macrobrachium nipponense and Eriocheir sinensis: Comparative analysis of structure and activities

Qian

Yang

et al. 2012

Fish & Shellfish Immunology

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Three Kazal-type serine proteinase inhibitors from the red swamp crayfish Procambarus clarkii and the characterization, function analysis of hcPcSPI2

Cited by 27 publications

References 35 publications

Identification, Characterization, and Functional Analysis of Tube and Pelle Homologs in the Mud Crab Scylla paramamosain

Identification, Characterization, and Functional Analysis of Tube and Pelle Homologs in the Mud Crab Scylla paramamosain

Effects of enrofloxacin on cytochromes P4501A and P4503A in Carassius auratus gibelio (crucian carp)

Two Kazal-type protease inhibitors from Macrobrachium nipponense and Eriocheir sinensis: Comparative analysis of structure and activities

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