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首页> 外文期刊>Journal of Cellular Physiology >Constitutive secretion of beta-trace protein by cultivated porcine choroid plexus epithelial cells: elucidation of its complete amino acid and cDNA sequences.
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Constitutive secretion of beta-trace protein by cultivated porcine choroid plexus epithelial cells: elucidation of its complete amino acid and cDNA sequences.

机译：本构beta-trace蛋白质的分泌培养猪脉络丛上皮细胞:完整的氨基酸和说明互补脱氧核糖核酸序列。

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摘要

Primary porcine choroid plexus epithelial cells cultivated in chemically defined medium maintain their epithelial characteristics and form confluent monolayers. They produce a fluid the composition of which resembles cerebrospinal fluid. The present study demonstrates constitutive secretion of large amounts of beta-trace protein. This intrathecally synthesized protein is a prominent polypeptide constituent of natural cerebrospinal fluid. According to the identity of amino acid sequences it has previously been tentatively identified as a prostaglandin-D synthase and as a member of the lipocalin protein family. beta-Trace was purified from cell culture supernatants and was subjected to tryptic digestion and amino acid sequencing of the resulting peptides. The complete primary structure of the protein was obtained by additional isolation of the cDNA from cultured epithelial cells. The porcine 163-amino acid polypeptide showed 69% identity with the human beta-trace and contained two N-glycosylation sites occupied by complex-type oligosaccharides as is the case for the human protein. The amino acid sequences around the N-glycosylation sites of mammalian beta-trace proteins (porcine, human, murine, and rat) were highly conserved. The nucleotide sequence was found to be less conserved; the porcine cDNA had a strikingly high GC-content (67%). The constitutive secretion of beta-trace protein from the in vitro cultivated porcine choroid plexus epithelial cells demonstrates that the cells have retained their major in vivo physiological properties: secretion of cerebrospinal fluid proteins. Therefore, this in vitro culture system may be used as a versatile tool for studying the regulation of the formation of cerebrospinal fluid.
机译：主要猪脉络丛上皮细胞培养化学定义中维护他们的上皮和形式特征汇合的单层膜。类似于脑脊髓的成分液体。本构分泌大量的beta-trace蛋白质。是一个杰出的多肽合成蛋白质组成的自然脑脊液。根据氨基酸序列的身份此前已经初步确认prostaglandin-D合成酶和作为的一员lipocalin蛋白质家族。从细胞培养上清液和受到胰蛋白酶的消化和氨基酸序列由此产生的肽。获得的蛋白质的结构额外的隔离的互补的培养上皮细胞。多肽显示,69%的身份与人类beta-trace,包含两个N-glycosylation网站被复杂类型低聚糖一样的人类蛋白质。酸序列在N-glycosylation网站哺乳动物beta-trace蛋白质(猪、人类、小鼠和大鼠)是高度保守的。核苷酸序列被发现守恒的;GC-content(67%)。beta-trace蛋白的体外培养猪脉络丛上皮细胞表明细胞保留他们主要体内生理属性:分泌脑脊液的蛋白。体外培养系统可以用作多用途工具学习的规定脑脊液的形成。

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