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Isolation and Characterization of Adhesive Secretion fromCuvierian Tubules of Sea Cucumber Holothuria forskali (Echinodermata: Holothuroidea)

机译:海参库维氏小管的Cuvierian肾小管黏附分泌物的分离和鉴定(Echinodermata:Holothuroidea)

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@@ The sea cucumber Holothuria forsk°ali possesses a specialized systemcalled Cuvierian tubules. During mechanical stimulation white filaments (tubules) are expelled and become sticky upon contact with any object. We isolated a protein with adhesive properties from protein extracts of Cuvierian tubules from H. Forsk°ali. This protein was identified by antibodies against recombinant precollagen D which is located in the byssal threads of the mussel Mytilus galloprovincialis. To find out the optimal procedure for extraction and purification, the identified protein was isolated by several methods, including electroelution, binding to glass beads, immunoprecipitation, and gel filtration. Antibodies raised against the isolated protein were used for localization of the adhesive protein in Cuvierian tubules. Immunostaining and immunogold electron microscopical studies revealed the strongest immunoreactivity in themesothelium; this tissue layer is involved in adhesion. Adhesion of Cuvierian tubule extracts was measured on the surface of various materials. The extracted protein showed the strongest adhesion to Teflon surface. Increased adhesion was observed in the presence of potassium and EDTA, while cadmium caused a decrease in adhesion. Addition of antibodies and trypsin abolished the adhesive properties of the extract.
机译:@@海参Holothuria forsk°ali具有称为Cuvierian小管的专门系统。在机械刺激过程中,白色细丝(管状)被排出并在与任何物体接触时变得发粘。我们从H. Forsk°ali的Cuvierian小管蛋白提取物中分离出了具有粘附特性的蛋白。该蛋白是通过针对位于贻贝Mytilus galloprovincialis底脉中的重组前胶原D的抗体鉴定的。为了找到提取和纯化的最佳程序,可通过几种方法分离鉴定出的蛋白质,包括电洗脱,与玻璃珠的结合,免疫沉淀和凝胶过滤。针对分离的蛋白质产生的抗体被用于在Cuvierian小管中定位黏附蛋白。免疫染色和免疫金电子显微镜研究揭示了上皮细胞中最强的免疫反应性。该组织层参与粘附。在各种材料的表面上测量了Cuvierian小管提取物的粘附力。提取的蛋白质显示出对特氟隆表面的最强粘附力。在钾和EDTA的存在下,粘附力增加,而镉导致粘附力下降。抗体和胰蛋白酶的加入消除了提取物的粘附性。

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