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利用基质辅助激光解吸/电离源内衰变对小鼠睾丸中组蛋白H3变体赖氨酸9上的翻译后修饰进行表征。

Characterization of post-translational modifications on lysine 9 of histone H3 variants in mouse testis using matrix-assisted laser desorption/ionization in-source decay.

作者信息

Kwak Ho-Geun, Dohmae Naoshi

机构信息

Biomolecular Characterization Unit, RIKEN Center for Sustainable Resource Science, 2-1 Hirosawa, Wako, 351-0198, Japan.

Graduate School of Science and Engineering, Saitama University, Saitama, Saitama, 338-8570, Japan.

出版信息

Rapid Commun Mass Spectrom. 2016 Dec 15;30(23):2529-2536. doi: 10.1002/rcm.7742.

DOI:10.1002/rcm.7742
PMID:27643486
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5108415/
Abstract

RATIONALE

Post-translational modifications (PTMs) of histones result in changes to transcriptional activities and chromatin remodeling. Lysine 9 of histone H3 (H3K9) is subject to PTMs, such as methylation and acetylation, which influence histone activity during spermatogenesis. Characterization strategies for studying PTMs on H3K9 have been developed to provide epigenetic and proteomic information. Proteomic analysis has been used to limited success to study PTMs on H3K9; however, a comprehensive analytical approach is required to elucidate global patterns of PTMs of H3 variants during spermatogenesis.

METHODS

Intact H3 variants in mouse testis were separated by high-performance liquid chromatography on a reversed-phase column with an ion-pairing reagent. Modifications to H3K9 were identified via top-down analysis using matrix-assisted laser desorption/ionization in source decay (MALDI-ISD).

RESULTS

Mono-, di-, and tri-methylations were identified at H3K9 in mouse testis and epididymis. These modifications were also observed in testis-specific histone H3 (H3t). Specifically, tri-methylation was more abundant on H3tK9 than on K9 of other H3 variants.

CONCLUSIONS

We introduce a method for rapid, simple, and comprehensive characterization of PTMs on the N-termini of H3 variants using MALDI-ISD. This approach provides novel and useful information, including K9 modifications on H3t, which would benefit epigenetic and proteomic research. © 2016 The Authors. Rapid Communications in Mass Spectrometry Published by John Wiley & Sons Ltd.

摘要

原理

组蛋白的翻译后修饰（PTM）会导致转录活性和染色质重塑发生变化。组蛋白H3（H3K9）的赖氨酸9会发生PTM，如甲基化和乙酰化，这些修饰会影响精子发生过程中的组蛋白活性。已经开发出用于研究H3K9上PTM的表征策略，以提供表观遗传学和蛋白质组学信息。蛋白质组学分析在研究H3K9上的PTM方面取得了有限的成功；然而，需要一种全面的分析方法来阐明精子发生过程中H3变体PTM的全局模式。

方法

通过在反相柱上使用离子对试剂的高效液相色谱法分离小鼠睾丸中的完整H3变体。通过使用源内衰变基质辅助激光解吸/电离（MALDI-ISD）的自上而下分析来鉴定H3K9的修饰。

结果

在小鼠睾丸和附睾中的H3K9处鉴定出单甲基化、二甲基化和三甲基化。在睾丸特异性组蛋白H3（H3t）中也观察到了这些修饰。具体而言，H3tK9上的三甲基化比其他H3变体的K9上的三甲基化更丰富。

结论

我们介绍了一种使用MALDI-ISD对H3变体N端PTM进行快速、简单和全面表征的方法。这种方法提供了新颖且有用的信息，包括H3t上的K9修饰，这将有益于表观遗传学和蛋白质组学研究。©2016作者。《质谱快报》由约翰·威利父子有限公司出版。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/ee0f440fbe41/RCM-30-2529-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/cadb2b845da7/RCM-30-2529-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/69f47df44104/RCM-30-2529-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/f4fb6a58771c/RCM-30-2529-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/9740585f245a/RCM-30-2529-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/e473462e32b5/RCM-30-2529-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/9a70abdedba1/RCM-30-2529-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/ee0f440fbe41/RCM-30-2529-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/cadb2b845da7/RCM-30-2529-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/69f47df44104/RCM-30-2529-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/f4fb6a58771c/RCM-30-2529-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/9740585f245a/RCM-30-2529-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/e473462e32b5/RCM-30-2529-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/9a70abdedba1/RCM-30-2529-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c228/5108415/ee0f440fbe41/RCM-30-2529-g007.jpg

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2
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Reproduction. 2016 May;151(5):R55-70. doi: 10.1530/REP-15-0562. Epub 2016 Feb 5.
3
Tissue-specific expression of histone H3 variants diversified after species separation.
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Epigenetics Chromatin. 2015 Sep 17;8:35. doi: 10.1186/s13072-015-0027-3. eCollection 2015.
4
MALDI-ISD Mass Spectrometry Analysis of Hemoglobin Variants: a Top-Down Approach to the Characterization of Hemoglobinopathies.血红蛋白变体的基质辅助激光解吸电离离子淌度质谱分析：一种自上而下表征血红蛋白病的方法。
J Am Soc Mass Spectrom. 2015 Aug;26(8):1299-310. doi: 10.1007/s13361-015-1164-4. Epub 2015 May 22.
5
Top-down MALDI-in-source decay-FTICR mass spectrometry of isotopically resolved proteins.同位素分辨蛋白质的自上而下的基质辅助激光解吸电离源内衰变-傅里叶变换离子回旋共振质谱分析
Anal Chem. 2015 Mar 17;87(6):3429-37. doi: 10.1021/ac504708y. Epub 2015 Mar 5.
6
Principles of hydrogen radical mediated peptide/protein fragmentation during matrix-assisted laser desorption/ionization mass spectrometry.基质辅助激光解吸电离质谱中氢自由基介导的肽/蛋白质片段化原理。
Mass Spectrom Rev. 2016 Jul;35(4):535-56. doi: 10.1002/mas.21444. Epub 2014 Oct 6.
7
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