Analytical Chemistry

Mapping the Proteoform Landscape of Five Human Tissues

Authors

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 human lung, heart, spleen, small intestine, and kidney. CZE and RPLC provided complementary post-translational modification (PTM) and proteoform selectivity, thereby enhancing overall proteome coverage when used in combination. Of the 11,466 proteoforms identified in this study, 7,373 (64%) were not reported previously. Large differences in 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.

Content

Thumbnail image of ChemRxiv_HuBMAP_Tissues_v1.pdf

Supplementary material

Thumbnail image of Supplementary_Data_1_Unique_PFRs.xlsx
List of tissue-specific proteoform identified in this study
Proteoforms were identified by top-down proteomics. Highly characterized proteoforms that were observed in only one tissue type are listed here.

Supplementary weblinks

Human Proteoform Atlas
Proteoforms identified in this study were deposited into the Human Proteoform Atlas