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.
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.
Hsieh
S
S
Department of Molecules and Cellular Neurobiology, Graduate School Neurosciences Amsterdam, Vrije Universiteit, Faculty of Biology, The Netherlands.
Dreisewerd
K
K
van der Schors
R C
RC
Jiménez
C R
CR
Stahl-Zeng
J
J
Hillenkamp
F
F
Jorgenson
J W
JW
Geraerts
W P
WP
Li
K W
KW
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
D009479:Neuropeptides; D010446:Peptide Fragments