کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8978130 | 1106574 | 2005 | 16 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Molecular characterization of a novel pattern recognition protein from nonspecific cytotoxic cells: Sequence analysis, phylogenetic comparisons and anti-microbial activity of a recombinant homologue
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شناسی تکاملی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Molecular characterization of a novel pattern recognition protein from nonspecific cytotoxic cells: Sequence analysis, phylogenetic comparisons and anti-microbial activity of a recombinant homologue Molecular characterization of a novel pattern recognition protein from nonspecific cytotoxic cells: Sequence analysis, phylogenetic comparisons and anti-microbial activity of a recombinant homologue](/preview/png/8978130.png)
چکیده انگلیسی
Nonspecific cytotoxic cells (NCC) are the first identified and most extensively studied killer cell population in teleosts. NCC kill a wide variety of target cells including tumor cells, virally transformed cells and protozoan parasites. The present study identified a novel evolutionarily conserved oligodeoxynucleotide (ODN) binding membrane protein expressed by channel catfish (Ictalurus punctatus) NCC. Peptide fingerprinting analysis of the ODN binding protein (referred to as NCC cationic anti-microbial protein-1/ncamp-1) identified a peptide that was used to design degenerate primers. A catfish NCC cDNA library was used as template with these primers and the PCR-amplified product was sequenced. The translated sequence contained 203 amino acids (molecular mass of 22,064.63Â Da) with characteristic lysine rich regions and a pI=pH 10.75. Sequence comparisons of this protein indicated similarity to zebrafish (51.2%) histone family member 1-X and (to a lesser extent) to trout H1. A search of EST databases confirmed that ncamp-1 is also expressed in various tissues of channel catfish as well as zebrafish. Inspection for signature repeats in ncamp-1 and comparisons with histone-like peptides from different species indicated the presence of multiple lysine based motifs composed of AKKA or PKK repeats. The novel protein was cloned, expressed in E. coli and the recombinant was used to generate rabbit anti-serum. The recombinant ncamp-1 bound GpC and CpG ODNs and was detected with homologous anti-ncamp-1 polyclonal antibodies. Western blots of NCC membranes using anti-ncamp-1 serum detected a 29Â kDa protein. Binding competition experiments demonstrated that anti-ncamp-1 antibodies and GpC bound to the same protein on NCC. Two different truncated forms of ncamp-1 as well as the full-length recombinant protein exhibited anti-microbial activity. The present study demonstrated the expression by NCC of a new membrane protein that may participate in the recognition of bacterial DNA and as such participate in innate anti-microbial immune responses in teleosts.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Developmental & Comparative Immunology - Volume 29, Issue 12, 2005, Pages 1049-1064
Journal: Developmental & Comparative Immunology - Volume 29, Issue 12, 2005, Pages 1049-1064
نویسندگان
Donald L. Evans, Harjeet Kaur, John III, Kesavannair Praveen, Liliana Jaso-Friedmann,