A C-type lectin, Nattectin-like protein (CaNTC) in Qihe crucian carp Carassius auratus: Binding ability with LPS, PGN and various bacteria, and agglutinating activity against bacteria. Issue 67 (August 2017)

• Record Type: Journal Article
• Authors: Wang, Li; Zhang, Jie; Kong, Xianghui; Zhao, Xianliang; Pei, Chao; Li, Li
• Is Part Of: Fish & shellfish immunology. Issue 67(2017)
• Page Start: 382
• Page End: 392
• Publication Date: 2017-08
• Subjects: C-type lectin -- Nattectin-like protein -- Bacterial-binding ability -- Agglutinating activity -- Innate immunity -- Carassius auratus; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2017.06.012
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: C-type lectins (CTLs), as the members of pattern-recognition receptors (PRRs), play the significant roles in innate immunity through binding with pathogen-associated molecular patterns (PAMPs) on the surface of microbe. In the present study, a novel CTL, Nattectin-like protein (named as CaNTC), was investigated in Qihe crucian carp Carassius auratus. The full-length cDNA of CaNTC was composed of 776 bp, with a 152 bp 5′-untranslated region (UTR), a 492 bp ORF encoding a 163-aa protein, and a 132 bp 3′-UTR with a polyadenylation signal sequence AATAAA and a poly(A) tail. The deduced amino acid sequence of CaNTC contained a signal peptide, a single carbohydrate recognition domain (CRD) which had four conserved disulfide-bonded cysteine residues (Cys 57 -Cys 150, Cys 126 -Cys 142 ), and an EPN/WND motif required for carbohydrate-binding specificity. With regard to the mRNA transcript of CaNTC, it was predominately expressed in liver. The temporal expressions of CaNTC were obviously up-regulated in liver, spleen and head-kidney after challenged by Aeromonas hydrophila and poly I: C, respectively, and the change pattern was in the time-depended manner. The recombinant CaNTC (rCaNTC) purified from Escherichia coli BL21 (DE3), exhibited strong binding ability with LPS and PGN, as well as all tested bacteria in a Ca 2+ -independent manner. With regard to the agglutinating activity of rCaNTC, rCaNTC was able to agglutinate rabbit erythrocytes and three kinds of bacteria (Gram-negative bacteria, Escherichia coli and A. hydrophila, and Gram-positive bacteria Staphylococcus aureus ) in a Ca 2+ -dependent manner. These findings collectively demonstrated that CaNTC, as a PRR, could be involved in the innate immunity and play an important role in immune defense of C. auratus . Highlights: A novel C-type lectin CaNTC was characterized in Qihe crucian carp Carassius auratus. CaNTC mRNA expressions were obviously up-regulated after pathogen challenges. Recombinant CaNTC exhibited binding ability with LPS, PGN and various bacteria. Recombinant CaNTC could agglutinate rabbit erythrocytes and some bacteria.