关键词: AQP2 aquaporin‐2 collecting duct kidney next‐generation DNA sequencing phosphoproteomics protein kinase A protein mass spectrometry proteomics vasopressin

Mesh : Aquaporin 2 / metabolism Proteomics / methods Animals Humans Vasopressins / metabolism Protein Processing, Post-Translational

来  源:   DOI:10.1113/JP283899   PDF(Pubmed)

Abstract:
The advent of modern quantitative protein mass spectrometry techniques around the turn of the 21st century has contributed to a revolution in biology referred to as \'systems biology\'. These methods allow identification and quantification of thousands of proteins in a biological specimen, as well as detection and quantification of post-translational protein modifications including phosphorylation. Here, we discuss these methodologies and show how they can be applied to understand the effects of the peptide hormone vasopressin to regulate the molecular water channel aquaporin-2. The emerging picture provides a detailed framework for understanding the molecular mechanisms involved in water balance disorders.
摘要:
在21世纪初,现代定量蛋白质质谱技术的出现促成了被称为“系统生物学”的生物学革命。这些方法可以识别和定量生物样本中的数千种蛋白质,以及包括磷酸化在内的翻译后蛋白质修饰的检测和定量。这里,我们讨论了这些方法,并展示了如何应用它们来了解肽激素加压素调节分子水通道水通道蛋白2(AQP2)的作用。新出现的图片提供了一个详细的框架,以了解水平衡障碍涉及的分子机制。摘要图例各种定量蛋白质质谱技术已被用于研究血管加压素诱导AQP2蛋白表达的机制。SILAC是一种使用同位素(C-13和N-15)标记细胞培养物中的精氨酸和赖氨酸的技术,该技术允许质谱法区分对照和处理样品。通过测量对应于样品中亲本蛋白质的相对丰度的轻肽和重肽的相对强度来进行定量。动态SILAC旨在通过将正常培养基切换到含有重同位素的培养基来量化蛋白质半衰期和翻译速率,并追踪轻肽和重肽的强度变化。TMT是一组具有相同分子量的同位素标签,可以直接标记肽。TMT技术的优点是多路复用,可以应用于任何样品类型,而无需与重同位素培养基进行预孵育。TMT定量的多路复用对于研究时间响应性磷酸蛋白质组学至关重要。本文受版权保护。保留所有权利。
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