title: Integration of surface plasmon resonance with mass spectrometry: automated ligand fishing and sample preparation for MALDI MS using a Biacore 3000 biosensor.
abstract: Integrating surface plasmon resonance analysis with mass spectrometry allows detection and characterization of molecular interactions to be complemented with identification of interaction partners. We have developed a procedure for Biacore 3000 that automatically performs all steps from ligand fishing and recovery to sample preparation for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry including on-target digestion. In the model system used in this study a signal transduction protein, calmodulin, was selectively captured from brain extract by one of its interaction partners immobilized on a sensor chip. The bound material was eluted, deposited directly onto a MALDI target, and analyzed by mass spectrometry both as an intact protein and after on-target tryptic digestion. The procedure with direct deposition of recovered material on the MALDI target reduces sample losses and, in combination with automatic sample processing, increases the throughput of surface plasmon resonance mass spectrometry analysis.
authors.affiliation: Biacore AB, Rapsgatan 7, 75450 Uppsala, Sweden.
authors.forename: Andrei
authors.initials: A
authors.lastname: Zhukov
authors.forename: Martin
authors.initials: M
authors.lastname: Schürenberg
authors.forename: Osten
authors.initials: O
authors.lastname: Jansson
authors.forename: Daphne
authors.initials: D
authors.lastname: Areskoug
authors.forename: Jos
authors.initials: J
authors.lastname: Buijs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D001921:Brain / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002417:Cattle; D030562:Databases, Protein; D008024:Ligands; D008969:Molecular Sequence Data; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D020539:Sequence Analysis, Protein / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D008024:Ligands; D010446:Peptide Fragments; D009219:Myosin-Light-Chain Kinase