Dataset Information

Integrated bottom-up and top-down proteomics of patient-derived breast tumor xenografts

ABSTRACT:

Ntai, I., LeDuc, R.D., Fellers, R.T., et al., (2015) Molecular and Cellular Proteomics, Manuscript M114.047480

Bottom-up proteomics relies on the use of proteases and is the method of choice for identifying thousands of protein groups in complex samples. Top-down proteomics has been shown to be robust for direct analysis of small proteins and offers a solution to the peptide-to- protein inference problem inherent with bottom-up approaches. Here, we describe the first large-scale integration of genomic, bottom-up and top-down proteomic data for the comparative analysis of patient-derived mouse xenograft models of basal and luminal B human breast cancer, WHIM2 and WHIM16, respectively. Using these well-characterized xenograft models established by the National Cancer Institute's Clinical Proteomic Tumor Analysis Consortium, we compared and contrasted the performance of bottom-up and top-down proteomics to detect cancer-specific aberrations at the peptide and proteoform levels, and to measure differential expression of proteins and proteoforms. Bottom-up proteomic analysis of the tumor xenografts detected almost 10 times as many coding nucleotide polymorphisms and peptides resulting from novel splice junctions than top-down. For proteins in the range of 0-30 kDa, where quantitation was performed using both approaches, bottom-up proteomics quantified 3,519 protein groups from 49,185 peptides, while top-down proteomics quantified 982 proteoforms mapping to 358 proteins. Examples of both concordant and discordant quantitation were found in an approximately 60:40 ratio, providing a unique opportunity for top-down to fill in missing information. The two techniques showed complementary performance, with bottom-up yielding 8 times more identifications of 0-30 kDa proteins in xenograft proteomes, but failing to detect differences in certain post-translational modifications (PTMs), such as phosphorylation pattern changes of alpha-endosulfine. This work illustrates the potency of a combined bottom-up and top-down proteomics approach to deepen our knowledge of cancer biology, especially when genomic data are available.

Dataset imported into MassIVE from https://cptac-data-portal.georgetown.edu/cptac/s/S028 on 08/28/19

INSTRUMENT(S): TripleTOF 5600, LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Mus Musculus (ncbitaxon:10090)

SUBMITTER: Neil L Kelleher

PROVIDER: MSV000084247 | MassIVE | Wed Aug 28 15:26:00 BST 2019

SECONDARY ACCESSION(S): PXD015909

REPOSITORIES: MassIVE

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Publications

Integrated Bottom-Up and Top-Down Proteomics of Patient-Derived Breast Tumor Xenografts.

Ntai Ioanna I LeDuc Richard D RD Fellers Ryan T RT Erdmann-Gilmore Petra P Davies Sherri R SR Rumsey Jeanne J Early Bryan P BP Thomas Paul M PM Li Shunqiang S Compton Philip D PD Ellis Matthew J C MJ Ruggles Kelly V KV Fenyö David D Boja Emily S ES Rodriguez Henry H Townsend R Reid RR Kelleher Neil L NL

Molecular & cellular proteomics : MCP 20151026 1

PMID: 26503891

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