Characterization of Chlamydomonas reinhardtii Core Histones by Top-Down Mass Spectrometry Reveals Unique Algae-Specific Variants and Post-Translational Modifications,Journal of Proteome Research

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Characterization of Chlamydomonas reinhardtii Core Histones by Top-Down Mass Spectrometry Reveals Unique Algae-Specific Variants and Post-Translational Modifications
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2017-12-08 00:00:00 , DOI: 10.1021/acs.jproteome.7b00780
Aliyya Khan ₁ , Carlo K. Eikani ₁ , Hana Khan ₁ , Anthony T. Iavarone ₂ , James J. Pesavento ₁

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The unicellular microalga Chlamydomonas reinhardtii has played an instrumental role in the development of many new fields (bioproducts, biofuels, etc.) as well as the advancement of basic science (photosynthetic apparati, flagellar function, etc.). Chlamydomonas’ versatility ultimately derives from the genes encoded in its genome and the way that the expression of these genes is regulated, which is largely influenced by a family of DNA binding proteins called histones. We characterize C. reinhardtii core histones, both variants and their post-translational modifications, by chromatographic separation, followed by top-down mass spectrometry (TDMS). Because TDMS has not been previously used to study Chlamydomonas proteins, we show rampant artifactual protein oxidation using established nuclei purification and histone extraction methods. After addressing oxidation, both histones H3 and H4 are found to each have a single polypeptide sequence that is minimally acetylated and methylated. Surprisingly, we uncover a novel monomethylation at lysine 79 on histone H4 present on all observed molecules. Histone H2B and H2A are found to have two and three variants, respectively, and both are minimally modified. This study provides an updated assessment of the core histone proteins in the green alga C. reinhardtii by top-down mass spectrometry and lays the foundation for further investigation of these essential proteins.

中文翻译：

自上而下的质谱表征莱茵衣藻核心组蛋白揭示了独特的藻类特定变体和翻译后的修改。

单细胞微藻莱茵衣藻在许多新领域（生物产品，生物燃料等）的发展以及基础科学（光合作用，鞭毛功能等）的发展中发挥了重要作用。衣藻的多功能性最终来自其基因组中编码的基因以及这些基因表达的调控方式，这在很大程度上受称为组蛋白的DNA结合蛋白家族的影响。我们表征C. reinhardtii核心组蛋白，这两个变体和它们的翻译后修饰，通过色谱分离，然后自上而下的质谱分析（TDMS）。因为之前尚未使用TDMS研究衣藻蛋白质，我们显示了使用已建立的核纯化和组蛋白提取方法的猖artifact的人为蛋白质氧化。解决氧化问题后，发现组蛋白H3和H4均具有最小化乙酰化和甲基化的单个多肽序列。出人意料的是，我们发现了所有观察到的分子上存在的组蛋白H4上赖氨酸79处的新型单甲基化。发现组蛋白H2B和H2A分别具有两个和三个变体，并且都进行了最小程度的修改。这项研究通过自上而下的质谱分析提供了对绿藻莱茵衣藻中核心组蛋白的最新评估，并为进一步研究这些必需蛋白奠定了基础。

更新日期：2017-12-10

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