Label-Based Mass Spectrometry Approaches for Robust Quantification of the Phosphoproteome and Total Proteome in Toxoplasma gondii

Label-Based Mass Spectrometry Approaches for Robust Quantification of the Phosphoproteome and Total Proteome in Toxoplasma gondii
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DOI:
10.1007/978-1-4939-9857-9_23
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发表时间:
2020-01-01
期刊:
TOXOPLASMA GONDII: METHODS AND PROTOCOLS
影响因子:
--
通讯作者:
Treeck, Moritz
Treeck, Moritz
中科院分区:
其他
文献类型:
--
作者:
Broncel, Malgorzata;Treeck, Moritz

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Protein phosphorylation plays a key role in regulating biological processes. Over 30% of the proteome is phosphorylated in most organisms and unraveling the function of the kinases that mediate these phosphorylation events requires the technology to reliably measure phosphorylation on proteins under various conditions. Advances in mass-spectrometry instrumentation, sample preparation, and labeling technologies now offer a range of quantification methods, each with their advantages and disadvantages. Here we describe in detail two different quantification methods, that is, stable isotope labeling by amino acids in cell culture and tandem mass tagging, combined with phosphopeptide enrichment strategies to measure the phosphoproteome of Toxoplasma parasites.