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首页> 外文期刊>Fish & Shellfish Immunology >Purification, characterization and functional analysis of a novel beta-1, 3-glucan binding protein from green tiger shrimp Penaeus semisulcatus
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Purification, characterization and functional analysis of a novel beta-1, 3-glucan binding protein from green tiger shrimp Penaeus semisulcatus

机译：绿虎虾半对虾对虾新型β-1、3-葡聚糖结合蛋白的纯化，鉴定和功能分析

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摘要

A beta-1, 3-Glucan binding protein (beta-GBP) was isolated from green tiger shrimp Penaeus semisulcatus and purified using laminarin precipitation and affinity chromatography on laminarin-Sepharose 6B column respectively. P. semisulcatus beta-GBP exhibits a single band with a molecular weight of 112 kDa on SDS-PAGE and pI of 5.9 in isoelectric focusing (IEF). Negative staining of P. semisulcatus beta-GBP showed large aggregates with crystalline surface when viewed by Electron Microscopy. Circular dichroism spectra of P. semisulcatus beta-GBP showed broad negative minimum wavelength extending from 200 to 250 nm can be attributed to the presence of beta-sheets in its secondary structure. P. semisulcatus beta-GBP comprises the specific binding affinity with the polysaccharide beta-1, 3-glucans (laminarin), this recognition and binding leads to the activation of prophenoloxidase cascade. Interestingly, P. semisulcatus beta-GBP also involved in the agglutination of baker's yeast, bacteria, erythrocytes (RBCs) and enhances the PO activity. Herein, we have investigated the importance of beta-GBP in innate immune response of P. semisulcatus and they implicate the evolutionary link with similar proteins found in other invertebrates. (C) 2013 Elsevier Ltd. All rights reserved.
机译：从绿虎虾半对虾Penaeus semisulcatus中分离出β-1、3-葡聚糖结合蛋白（β-GBP），并分别使用层板蛋白沉淀和亲和色谱在层板蛋白-琼脂糖6B柱上纯化。半硫化P-GBP在SDS-PAGE上显示一条分子量为112 kDa的单条带，等电聚焦（IEF）的pI为5.9。用电子显微镜观察时，半硫化脲β-GBP的阴性染色显示大的聚集体具有晶体表面。半硫化β-GBP的圆二色性光谱显示，从200到250 nm的较宽的最小负波长可归因于其二级结构中存在β-折叠。半硫化疟原虫β-GBP具有与多糖β-1、3-葡聚糖（laminarin）的特异性结合亲和力，这种识别和结合导致原酚氧化酶级联反应的激活。有趣的是，半硫化毕赤酵母β-GBP还参与了面包酵母，细菌，红细胞（RBC）的凝集并增强了PO的活性。在本文中，我们研究了β-GBP在半硫化假单胞菌固有免疫反应中的重要性，并且它们暗示了与在其他无脊椎动物中发现的相似蛋白质的进化联系。 （C）2013 Elsevier Ltd.保留所有权利。

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