Direct identification of proteins from T47D cells and murine brain tissue by matrix-assisted laser desorption/ionization post-source decay/collision-induced dissociation.
The purpose of this study is to determine the feasibility of the direct matrix-assisted laser desorption/ionization (MALDI) identification of proteins in fixed T47D breast cancer cells and murine brain tissues. The ability to identify proteins from cells and tissue may lead to biomarkers that effectively predict the onset of defined disease states, and their dynamic behavior could be an important hint for drug target discoveries. Direct tissue application of trypsin allows protein identification in cells and tissues, while maintaining spatial integrity and intracellular organization. Using a chemical printer, matrix was co-registered on trypsinized human T47D breast cancer cells and cryo-preserved sections of murine brain tissue, followed by MALDI post-source decay (PSD) or MALDI collision-induced dissociation (CID), respectively. Mass-to-charge (m/z) data from the cells and brain tissues were processed using Mascot software interrogation of the National Center for Biotechnology Information (NCBI) database. Histone H2B was identified from cultured T47D human breast cancer cells. Tubulin beta2 was identified from mouse brain cortex following an induced stroke. These results suggest that MALDI PSD/CID, combined with bioinformatics, can be used for the direct identification of proteins from cells and tissues. Refinements in preparation techniques may improve this approach to provide a tool for quantitative proteomics and clinical analysis.
Pevsner
Paul H
PH
Department of Pharmacology, New York University School of Medicine, New York, NY 10016, USA. paul.pevsner@med.nyu.edu
Naftolin
Frederick
F
Hillman
Dean E
DE
Miller
Douglas C
DC
Fadiel
Ahmed
A
Kogus
Alexander
A
Stern
Arnold
A
Samuels
Herbert H
HH
D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis / Q000737:chemistry; D001921:Brain / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D051379:Mice; D009363:Neoplasm Proteins / Q000032:analysis / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry*; D010449:Peptide Mapping / Q000379:methods*; D010865:Pilot Projects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D020521:Stroke / Q000378:metabolism*
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins