天冬酰胺 614 决定小鼠抗衰老蛋白 Klotho 的运输和功能

IF 5.1 2区 生物学 Q2 CELL BIOLOGY
Cells Pub Date : 2024-10-21 DOI:10.3390/cells13201743
Zahra Fanaei-Kahrani, Christoph Kaether
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引用次数: 0

摘要

Klotho 是一种抗衰老蛋白,其缺失会显著缩短小鼠的寿命,而过度表达则会延长寿命。Klotho 是一种 I 型跨膜蛋白,在其外显子结构域的八个位置有 N-糖基化。我们的研究表明,在小鼠 Klotho 中,N614 位(而不是 N161、N285 或 N346 位)的 N-糖基化或突变是 Klotho 从内质网(ER)转运出去的关键。因此,野生型 Klotho-EGFP 以及 N-糖基化突变体 N161Q、N285Q 和 N346Q 存在于质膜(PM),而 N614Q Klotho-EGFP 仅有少量存在于质膜,大部分蛋白质积聚在 ER 中。Klotho-EGFP N614Q 的蛋白质相互作用组分析表明,它与蛋白酶体相关蛋白质和参与ER蛋白质加工的蛋白质(如热休克蛋白和蛋白质二硫异构酶)的相互作用增加,表明蛋白质折叠受损。共免疫沉淀实验证实了 Klotho-EGFP N614Q 与 ER伴侣的相互作用。有趣的是,尽管 Klotho-EGFP N614Q 在 PM 中的含量很低,但在 FGF23 的存在下,它能有效地诱导 FGF 受体介导的 ERK 激活,这凸显了它在触发下游信号转导方面的功效,即使在 PM 中的含量有限。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Asparagine614 Determines the Transport and Function of the Murine Anti-Aging Protein Klotho.

Klotho is an anti-aging protein whose deletion significantly reduces lifespan in mice, while its over-expression increases lifespan. Klotho is a type-I transmembrane protein that is N-glycosylated at eight positions within its ectodomain. Our study demonstrates that N-glycosylation or mutation at position N614, but not at N161, N285, or N346 in mouse Klotho, is critically involved in the transport of Klotho out of the endoplasmic reticulum (ER). Consequently, while wild-type Klotho-EGFP as well as the N-glycosylation mutants N161Q, N285Q, and N346Q were present at the plasma membrane (PM), only small amounts of the N614Q Klotho-EGFP were present at the PM, with most of the protein accumulating in the ER. Protein interactome analysis of Klotho-EGFP N614Q revealed increased interactions with proteasome-related proteins and proteins involved in ER protein processing, like heat shock proteins and protein disulfide isomerases, indicative of impaired protein folding. Co-immunoprecipitation experiments confirmed the interaction of Klotho-EGFP N614Q with ER chaperons. Interestingly, despite the low amounts of Klotho-EGFP N614Q at the PM, it efficiently induced FGF receptor-mediated ERK activation in the presence of FGF23, highlighting its efficacy in triggering downstream signaling, even in limited quantities at the PM.

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来源期刊
Cells
Cells Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
9.90
自引率
5.00%
发文量
3472
审稿时长
16 days
期刊介绍: Cells (ISSN 2073-4409) is an international, peer-reviewed open access journal which provides an advanced forum for studies related to cell biology, molecular biology and biophysics. It publishes reviews, research articles, communications and technical notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided.
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