title: Separation and identification of peptides in single neurons by microcolumn liquid chromatography-matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and postsource decay analysis.
abstract: Microcolumn liquid chromatography (LC) was interfaced with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for separation and identification of peptides present in single neurons from the brain of the snail Lymnaea stagnalis. The nanoliter microcolumn LC effluent, mixed off-line with nanoliter matrix solution, was deposited onto the sample target every 60 s, producing fractions of approximately 145 nL in volume, which, upon drying, produced spots of approximately 1 mm in size. At the end of the chromatographic separation, fractions from the sample target were scanned by MALDI-TOF-MS. Identification of peptide peaks was achieved on the basis of LC elution order and mass information. Further identification based on sequence information was carried out for a native peptide fractionated by microcolumn LC from a single neuron with the postsource decay technique.
authors.affiliation: Department of Molecules and Cellular Neurobiology, Graduate School Neurosciences Amsterdam, Vrije Universiteit, Faculty of Biology, The Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Hsieh
authors.forename: K
authors.initials: K
authors.lastname: Dreisewerd
authors.forename: R C
authors.initials: RC
authors.lastname: van der Schors
authors.forename: C R
authors.initials: CR
authors.lastname: Jiménez
authors.forename: J
authors.initials: J
authors.lastname: Stahl-Zeng
authors.forename: F
authors.initials: F
authors.lastname: Hillenkamp
authors.forename: J W
authors.initials: JW
authors.lastname: Jorgenson
authors.forename: W P
authors.initials: WP
authors.lastname: Geraerts
authors.forename: K W
authors.initials: KW
authors.lastname: Li
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D008195:Lymnaea / Q000737:chemistry*; D008970:Molecular Weight; D009474:Neurons / Q000737:chemistry*; D009479:Neuropeptides / Q000032:analysis* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D009479:Neuropeptides; D010446:Peptide Fragments