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Surface‐Plasmon‐Resonance‐Based Chemical Proteomics: Efficient Specific Extraction and Semiquantitative Identification of Cyclic Nucleotide‐Binding Proteins from Cellular Lysates by Using a Combination of Surface Plasmon Resonance, Sequential Elution and Liquid Chromatography–Tandem Mass Spectrometry
Author(s) -
Visser Natasja F. C.,
Scholten Arjen,
van den Heuvel Robert H. H.,
Heck Albert J. R.
Publication year - 2007
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200600449
Subject(s) - surface plasmon resonance , proteomics , chemistry , tandem mass spectrometry , mass spectrometry , chromatography , small molecule , elution , quantitative proteomics , biochemistry , nanotechnology , materials science , nanoparticle , gene
Chemical proteomics is a powerful methodology for identifying the cellular targets of small molecules, however, it is biased towards abundant proteins. Therefore, quantitative strategies are needed to distinguish between specific and nonspecific interactions. Here, we explore the potential of the combination of surface plasmon resonance (SPR) coupled to liquid chromatography–tandem mass spectrometry (LC‐MS/MS) as an alternative approach in chemical proteomics. We coupled cGMP molecules to the SPR chip, and monitored the binding and dissociation of proteins from a human lysate by using sequential elution steps and SPR. The eluted proteins were subsequently identified by LC‐MS/MS. Our approach enabled the efficient and selective extraction of low‐abundant cyclic‐nucleotide‐binding proteins such as cGMP‐dependent protein kinase, and a quantitative assessment of the less‐ and nonspecific competitive binding proteins. The data show that SPR‐based chemical proteomics is a promising alternative for the efficient specific extraction and quantitative identification of small‐molecule‐binding proteins from complex mixtures.