Mapping the Proteoform Landscape of Five Human Tissues

Bryon S. Drown, Kevin Jooß, Rafael D. Melani, Cameron Lloyd-Jones, Jeannie M. Camarillo, Neil L. Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

A functional understanding of the human body requires structure-function studies of proteins at scale. The chemical structure of proteins is controlled at the transcriptional, translational, and post-translational levels, creating a variety of products with modulated functions within the cell. The term "proteoform" encapsulates this complexity at the level of chemical composition. Comprehensive mapping of the proteoform landscape in human tissues necessitates analytical techniques with increased sensitivity and depth of coverage. Here, we took a top-down proteomics approach, combining data generated using capillary zone electrophoresis (CZE) and nanoflow reversed-phase liquid chromatography (RPLC) hyphenated to mass spectrometry to identify and characterize proteoforms from the human lungs, heart, spleen, small intestine, and kidneys. CZE and RPLC provided complementary post-translational modification and proteoform selectivity, thereby enhancing the overall proteome coverage when used in combination. Of the 11,466 proteoforms identified in this study, 7373 (64%) were not reported previously. Large differences in the protein and proteoform level were readily quantified, with initial inferences about proteoform biology operative in the analyzed organs. Differential proteoform regulation of defensins, glutathione transferases, and sarcomeric proteins across tissues generate hypotheses about how they function and are regulated in human health and disease.

Original languageEnglish (US)
Pages (from-to)1299-1310
Number of pages12
JournalJournal of Proteome Research
Volume21
Issue number5
DOIs
StatePublished - May 6 2022

Funding

This material is based upon the work supported by the NIH Common Fund, through the Office of Strategic Coordination/Office of the Director under award UH3 CA246635 (N.L.K.), National Institute of General Medical Sciences of the National Institutes of Health grant P41 GM108569 (N.L.K.), and National Institute of Cancer of the National Institutes of Health grant F32 CA246894 (B.S.D.).

Keywords

  • capillary zone electrophoresis
  • heart
  • kidney
  • lung
  • proteomics
  • small intestine
  • spleen
  • top-down proteomics

ASJC Scopus subject areas

  • Biochemistry
  • General Chemistry

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