Dataset Information

Single shot CZE-MS/MS for phosphoproteomics

ABSTRACT: The dependence of capillary zone electrophoresis separations on the charge state of analyte is useful for the analysis of many post-translational modifications in proteins. In this work, we coupled CZE to an Orbitrap Fusion Lumos Tribrid platform with advanced peak determination algorithm for phosphoproteomics analysis. A linear polyacrylamide coated capillary with very low electroosmotic flow was used for the separation. The optimal injection volume is between 100 nL and 150 nL of phosphopeptides dissolved in 30 mM ammonium bicarbonate (pH 8.2) buffer, which produces a dynamic pH junction sample injection. Larger injection volumes resulted in serious peak broadening and decreased numbers of phosphopeptide identifications. The optimized system identified 4405 phosphopeptides from 220 ng of enriched phosphopeptides from mouse brain, which represents the state-of-the-art result for single shot CZE-ESI-MS/MS based phosphoproteome analysis.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive HF

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain

SUBMITTER: Zhenbin Zhang

LAB HEAD: Zhenbin Zhang

PROVIDER: PXD012888 | Pride | 2019-06-13

REPOSITORIES: Pride

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Single-Shot Capillary Zone Electrophoresis-Tandem Mass Spectrometry Produces over 4400 Phosphopeptide Identifications from a 220 ng Sample.

Zhang Zhenbin Z Hebert Alexander S AS Westphall Michael S MS Coon Joshua J JJ Dovichi Norman J NJ

Journal of proteome research 20190626 8

The dependence of capillary zone electrophoresis (CZE) separations on the charge state of the analyte is useful for the analysis of many post-translational modifications in proteins. In this work, we coupled CZE to an Orbitrap Fusion Lumos Tribrid platform with an advanced peak determination algorithm for phosphoproteomics analysis. A linear-polyacrylamide-coated capillary with very low electroosmotic flow was used for the separation. The optimal injection volume was between 100 and 150 nL of a ...[more]

PMID: 31180221

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			Action	DRS
	20180220004_mb_PHOS_1.msf	Msf
	20180220004_mb_PHOS_1.raw	Raw
	20180220005_mb_PHOS_2.msf	Msf
	20180220005_mb_PHOS_2.raw	Raw
	20180704001.RAW	Raw