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Analytical Chemistry

Proteomics goes native

Method allows native mass spectrometry to evaluate multitudes of proteins in their intact, biological forms

by Celia Henry Arnaud
November 20, 2017 | APPEARED IN VOLUME 95, ISSUE 46

When scientists analyze proteins with mass spectrometry, they typically denature the biomolecules, causing the proteins’ biologically relevant forms and composition to disappear. In contrast, so-called native mass spectrometry allows proteins and their complexes to remain intact during analysis.

For the past 15 years, though, native mass spectrometry has been relegated to analyzing specific, targeted protein complexes rather than unknown mixtures of them, says Neil L. Kelleher, a professor of chemistry at Northwestern University.

Now a team of researchers led by Kelleher and Northwestern’s Philip D. Compton has expanded native mass spectrometry into native proteomics, which they can use in an untargeted discovery mode to analyze endogenous complexes in cells and tissues (Nat. Chem. Biol. 2017, DOI: 10.1038/nchembio.2515). “This technique represents the next step in the evolution of native MS,” Kelleher says.

In their new method, the researchers extract protein complexes from cells and tissue, separate them, and analyze them by native top-down mass spectrometry. The native conditions preserve interactions that would be lost in harsher conditions. The team characterized 125 intact protein complexes and 217 distinct protein molecular species, including posttranslational modifications (PTMs), extracted from mouse heart and human cancer cell lines.

For example, the researchers observed a previously unreported Mg2+ binding site in a dimer of the human α-enolase enzyme. The binding site wasn’t present on the enzyme monomer. The team was also able to identify sequence variations and PTMs of cysteines on protein surfaces, which are often lost in denaturing proteomics.

The work is “impressive,” says Joseph A. Loo, a biochemistry professor at UCLA who also uses native mass spectrometry. “Their experimental platform contributes additional value to the typical proteomics output by adding information on ligand binding, PTMs, and functional assemblies.”

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Comments
Enrico Petrucco (November 20, 2017 6:55 AM)
This is very interesting. I hope this advancement in Proteomics will hopefully be quickly adopted to further the Cancer Genome Atlas.
Syed Azmal Ali (November 20, 2017 11:58 PM)
Very Exciting for the mass spectrometer based biological experimental designs. Looking forward to learn this new approach and provide some logical insights.

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