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Mass Spectrometry: Complex Analysis

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...

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Published on: May 17, 2016

Profiling Changes in Histone Post-translational Modifications by Top-Down Mass Spectrometry.

Mowei Zhou¹, Si Wu^1,2, David L Stenoien¹

¹Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|November 11, 2016

Summary

Top-down mass spectrometry (MS) rapidly profiles histone proteoforms and their modifications. This method enables comparative analysis of gene expression changes between different biological samples, like wild-type versus mutant species.

Keywords:

Histone Liquid chromatography Mass spectrometry Post-translational modification Screening Top-down

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Published on: January 12, 2024

Area of Science:

Biochemistry
Genomics
Proteomics

Background:

Gene expression is regulated by histone post-translational modifications, collectively known as the histone code.
Understanding the histone code is crucial for deciphering cellular functions and disease mechanisms.

Purpose of the Study:

To present a top-down mass spectrometry (MS) workflow for rapid, global profiling of histone proteoforms.
To demonstrate the utility of this workflow for comparative analysis of histone modifications between different biological states.

Main Methods:

A top-down mass spectrometry workflow utilizing liquid chromatography (LC) coupled to MS (LC-MS) was developed.
The method was applied to compare histone proteoforms between a wild-type and a mutant fungal species.

Main Results:

The LC-MS top-down approach enabled fast global profiling of histone proteoforms.
Differential abundances of specific proteoforms were identified between the wild-type and mutant fungal species.

Conclusions:

This top-down LC-MS method provides a powerful tool for analyzing the histone code and identifying changes in histone modifications.
The protocol is adaptable for screening histone modification alterations in various comparative studies, including wild-type vs. mutant or healthy vs. diseased samples.