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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

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Published on: November 15, 2017

High-Throughput Quantitative Proteomics Enabled by Mass Defect-Based 12-Plex DiLeu Isobaric Tags.

Dustin C Frost¹, Lingjun Li^2,3

¹School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI, 53705, USA.

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

|February 13, 2016

Summary

This summary is machine-generated.

Researchers developed cost-effective N,N-dimethyl leucine (DiLeu) isobaric tags for multiplexed mass spectrometry (MS) based proteomics. This method enables high-throughput relative quantification of up to twelve samples, overcoming cost barriers of commercial alternatives.

Keywords:

DiLeu High-resolution mass spectrometry Isobaric labeling Isobaric tag synthesis Mass defect Multiplexed quantitation Quantitative proteomics

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Area of Science:

Proteomics
Analytical Chemistry
Biochemistry

Background:

Isobaric labeling is crucial for high-throughput mass spectrometry (MS)-based relative quantification of peptides and proteins.
The high cost of commercial isobaric tags limits their application in large-scale proteomics.
A need exists for cost-effective and accessible isobaric labeling strategies.

Purpose of the Study:

To develop a cost-effective alternative to commercial isobaric tags for multiplexed quantitative proteomics.
To present detailed methods for synthesizing and utilizing N,N-dimethyl leucine (DiLeu) multiplex isobaric tags.
To demonstrate the application of DiLeu tags for high-resolution MS-based relative quantification.

Main Methods:

Synthesis of N,N-dimethyl leucine (DiLeu) multiplex isobaric tags using readily available isotopic reagents.
Labeling of complex protein digest samples with the developed DiLeu tags.
Analysis of labeled samples using high-resolution nano-liquid chromatography coupled with tandem mass spectrometry (nanoLC-MS(n)).

Main Results:

Successful synthesis of 12-plex DiLeu isobaric tags.
Demonstration of DiLeu tags enabling relative quantification of up to twelve samples in a single LC-MS(2) experiment.
Mass defect-based DiLeu tags are compatible with high-resolution MS(n) acquisition for accurate quantification.

Conclusions:

N,N-dimethyl leucine (DiLeu) multiplex isobaric tags offer a cost-effective solution for large-scale proteomics.
The developed DiLeu labeling strategy facilitates high-throughput, multiplexed relative quantification of proteins.
This approach enhances the accessibility of advanced quantitative proteomics techniques.