| NanoLC-FT-ICR MS improves proteome coverage attainable for approximately 3000 laser-microdissected breast carcinoma cells. | |
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MedLine Citation:
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PMID: 17163580 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Proteomics assays hold great promise for unraveling molecular events that underlie human diseases. Effective analysis of clinical samples is essential, but this task is considerably complicated by tissue heterogeneity. Laser capture microdissection (LCM) can be used to selectively isolate target cells from their native tissue environment. However, the small number of cells that is typically procured by LCM severely limits proteome coverage and biomarker discovery potential achievable by conventional proteomics platforms. Herein, we describe the use of nanoLC-FT-ICR MS for analyzing protein digests of 3000 LCM-derived tumor cells from breast carcinoma tissue, corresponding to 300 ng of total protein. A total of 2282 peptides were identified by matching LC-MS data to accurate mass and time (AMT) tag databases that were previously established for human breast (cancer) cell lines. One thousand and three unique proteins were confidently identified with two or more peptides. Based on gene ontology categorization, identified proteins appear to cover a wide variety of biological functions and cellular compartments. This work demonstrates that a substantial number of proteins can be detected and identified from limited number of cells using the AMT tag approach, and opens doors for high-throughput in-depth proteomics analysis of clinical samples. |
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Authors:
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Arzu Umar; Theo M Luider; John A Foekens; Ljiljana Pasa-Tolić |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Proteomics Volume: 7 ISSN: 1615-9853 ISO Abbreviation: Proteomics Publication Date: 2007 Jan |
Date Detail:
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Created Date: 2007-01-10 Completed Date: 2007-03-14 Revised Date: 2007-12-03 |
Medline Journal Info:
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Nlm Unique ID: 101092707 Medline TA: Proteomics Country: Germany |
Other Details:
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Languages: eng Pagination: 323-9 Citation Subset: IM |
Affiliation:
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Department of Medical Oncology, Erasmus Medical Center Rotterdam, Josephine Nefkens Institute, The Netherlands. a.umar@erasmusmc.nl |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Breast Neoplasms
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metabolism* Carcinoma / metabolism* Chromatography, Liquid Female Humans Lasers* Microdissection* Nanotechnology* Proteome / analysis*, metabolism Spectrometry, Mass, Electrospray Ionization* Spectroscopy, Fourier Transform Infrared* |
| Grant Support | |
ID/Acronym/Agency:
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RR18522/RR/NCRR NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Proteome |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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