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首页> 外文学位 >Molecular cloning and functional expression of an intestinal lactoferrin receptor and feasibility of using recombinant human lactoferrin expressed in rice in infant nutrition.
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Molecular cloning and functional expression of an intestinal lactoferrin receptor and feasibility of using recombinant human lactoferrin expressed in rice in infant nutrition.

机译：肠道乳铁蛋白受体的分子克隆和功能表达以及在婴儿营养中使用在水稻中表达的重组人乳铁蛋白的可行性。

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摘要

Lactoferrin (Lf), an 80 kD, single-chain glycoprotein, is abundant in human milk, binds iron tightly, and has been proposed to have many functions such as facilitating iron absorption, antimicrobial, and immunomodulating activities. Lf receptors (LfRs) have been found in several tissues including small intestine, liver, and lymphocytes, indicating that a primary biological function of Lf is mediated through these LfRs.; We here for the first time demonstrate the cloning of a UR gene from human and mouse small intestine, expression of the recombinant UR in a baculovirus-insect cell system, and its interaction with Lf. RT-PCR revealed that human UR is expressed at especially high levels in human fetal small intestine, but not as much in adult small intestine or in Caco-2 cells. The LfR is anchored to the apical membrane of Caco-2 cells by glycophosphatidyl inositol and uptake of Lf-bound 59Fe by LfR-transfected Caco-2 cells was significantly higher than that by mock-transfected cells. These results indicate that the UR on the apical membrane of human enterocytes mediates Lf-bound Fe transport. Mouse UR is expressed in early embryos, suggesting that the receptor-mediated function of Lf is important for embryonic development and that the mouse can be used to investigate biological functions of Lf in vivo. We also developed a recombinant HU (rHLf) expression system using rice and achieved a production level sufficient for commercial applications. We observed that rHLf has characteristics identical to native HU (nHLf) with regard to iron binding, antimicrobial activity, and binding/uptake to Caco-2 cells. Stability of rHLf after exposure to heat, low pH, or in-vitro digestion was also similar to nHLf, suggesting that rHLf can be used in infant foods.; In summary, we disclose genetic and biochemical information on the LfR, which is useful for studying the primary function of Lf Recombinant HLf expressed in rice is likely to act in infant small intestine in a manner similar to nHLf. Those functions include receptor-mediated mechanisms, possibly facilitating iron absorption and/or regulating the immune system, and non-receptor-mediated mechanisms such as antimicrobial activity.
机译：乳铁蛋白（Lf）是一种80 kD的单链糖蛋白，富含于人乳中，与铁紧密结合，并被提议具有许多功能，如促进铁吸收，抗菌和免疫调节活性。 Lf受体（LfRs）已在包括小肠，肝脏和淋巴细胞在内的多种组织中发现，表明Lf的主要生物学功能是通过这些LfRs介导的。我们在这里首次展示了从人和小鼠小肠克隆UR基因，在杆状病毒-昆虫细胞系统中重组UR的表达及其与Lf的相互作用。 RT-PCR显示，人UR在人胎儿小肠中的表达特别高，但在成年小肠或Caco-2细胞中表达不高。 LfR通过糖磷脂酰肌醇锚定在Caco-2细胞的顶膜上，LfR转染的Caco-2细胞对Lf结合的 59 Fe的摄取显着高于模拟转染的细胞。这些结果表明，人肠细胞顶膜上的UR介导Lf结合的Fe转运。小鼠UR在早期胚胎中表达，这表明Lf的受体介导的功能对于胚胎发育很重要，并且该小鼠可用于研究Lf的体内生物学功能。我们还开发了使用水稻的重组HU（rHLf）表达系统，并达到了足以用于商业用途的生产水平。我们观察到，rHLf在铁结合，抗菌活性以及对Caco-2细胞的结合/摄取方面具有与天然HU（nHLf）相同的特征。 rHLf暴露于热，低pH或体外消化后的稳定性也类似于nHLf，这表明rHLf可用于婴儿食品。总之，我们公开了有关LfR的遗传和生化信息，这对于研究Lf水稻中表达的重组HLf的主要功能很有用，它可能以类似于nHLf的方式作用于婴儿小肠。这些功能包括受体介导的机制，可能促进铁的吸收和/或调节免疫系统，以及非受体介导的机制，例如抗微生物活性。

著录项

  • 作者

    Suzuki, Yasushi A.;

  • 作者单位

    University of California, Davis.;

  • 授予单位 University of California, Davis.;
  • 学科 Health Sciences Nutrition.; Biology Molecular.
  • 学位 Ph.D.
  • 年度 2001
  • 页码 139 p.
  • 总页数 139
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 预防医学、卫生学;分子遗传学;
  • 关键词

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