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Originally published In Press as doi:10.1074/mcp.M600295-MCP200 on March 14, 2007.
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Molecular & Cellular Proteomics 6:1147-1157, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Identification of Glioma Neovascularization-related Proteins by Using MALDI-FTMS and Nano-LC Fractionation to Microdissected Tumor Vessels*

Dana A. N. Mustafa{ddagger}, Peter C. Burgers§, Lennard J. Dekker§, Halima Charif§, Mark K. Titulaer§, Peter A. E. Sillevis Smitt§, Theo M. Luider§,¶ and Johan M. Kros{ddagger},¶,||

From the {ddagger} Department of Pathology and § Laboratory of Neuro-oncology and Clinical Proteomics, Department of Neurology, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GD Rotterdam, The Netherlands

The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.

|| To whom correspondence should be addressed. Tel./Fax: 31-10-4087905; E-mail: j.m.kros{at}erasmusmc.nl


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