4.7 Article

Deep Proteome Mapping of Mouse Kidney Based on OFFGel Prefractionation Reveals Remarkable Protein Post- Translational Modifications

期刊

JOURNAL OF PROTEOME RESEARCH
卷 13, 期 3, 页码 1636-1646

出版社

AMER CHEMICAL SOC
DOI: 10.1021/pr401122m

关键词

OFFGel; kidney; proteomics; fractionation; phospho-enrichment

资金

  1. JSPS (Japan Society for Promotion of Science) [23790933, 21390262, 228071]
  2. Strategic young researcher overseas visits program for accelerating brain circulation [S2302]
  3. NIH [P4I GMI 03533, R01 MH067880, V19 AI063603]
  4. Grants-in-Aid for Scientific Research [23790933, 14F04105, 24591190, 25461212, 21390262] Funding Source: KAKEN

向作者/读者索取更多资源

Performing a comprehensive nonbiased proteome analysis is an extraordinary challenge due to sample complexity and wide dynamic range, especially in eukaryotic tissues. Thus, prefractionation steps conducted prior to mass spectrometric analysis are critically important to reduce complex biological matrices and allow in-depth analysis. Here we demonstrated the use of OFFGel prefractionation to identify more low abundant and hydrophobic proteins than in a nonfractionated sample. Moreover, OFFGel prefractionation of a kidney protein sample was able to unveil protein functional relevance by detecting PTMs, especially when prefractionation was augmented with a targeted enrichment strategy such as TiO2 phospho-enrichment. The OFFGel-TiO2 combination used in this study was comparable to other global phosphoproteomics approaches (SCX-TiO2 ERLIC-TiO2, or HILIC-TiO2). The detailed mouse kidney proteome with the phosphopeptide enrichment presented here serves as a useful platform for a better understanding of how the renal protein modification machinery works and, ultimately, will contribute to our understanding of pathological processes as well as normal physiological renal functions.

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