کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2431537 1106763 2013 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Isolation and characterization of a c-type lysozyme from the nurse shark
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم آبزیان
پیش نمایش صفحه اول مقاله
Isolation and characterization of a c-type lysozyme from the nurse shark
چکیده انگلیسی


• Characterization of shark lysozyme from unstimulated leukocytes.
• Active site sequence and Western blot reactivity shows it is a c-type lysozyme.
• Two isoforms of lysozyme are present in the shark.
• For LC-MS/MS analysis lysozyme can be isolated by one-step chromatography.

Lysozyme is a ubiquitous antibacterial enzyme that occurs in numerous invertebrate and vertebrate species. Three forms have been described c-type, g-type and i-type which differ in primary structure. Shark lysozyme has not been characterized; here we report on the isolation and characterization of lysozyme from unstimulated shark (Ginglymostoma cirratum) leukocytes and provide amino acid sequence data across the highly conserved active site of the molecule identifying it to be a c-type lysozyme. A leukocyte lysate was applied either (a) to the first of two sequential DE-52 cellulose columns or alternatively, (b) to a DEAE-Sepharose column. Lysozyme activity in lysate and active fractions was identified by zones of lysis of Micrococcus lysodeikticus cell walls on lysoplates and zones of growth inhibition in agar diffusion assays using Planococcus citreus as the target organism. SDS-PAGE analysis revealed a 14 kDa protein which was identified as lysozyme by mass spectroscopic analysis of peptides, reactivity against anti-HEWL antibodies on a Western blot, hydrolysis of M. lysodeikticus cell walls, and inhibition of growth of P. citreus on AU-gel blots in which the area of growth inhibition correlated to a 14 kDa protein.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Fish & Shellfish Immunology - Volume 35, Issue 6, December 2013, Pages 1824–1828
نویسندگان
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