Identification of POMC processing products in single melanotrope cells by matrix-assisted laser desorption/ionization mass spectrometry.
The use of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) in identifying proopiomelanocortin (POMC) processing products in melanotrope cells of the pituitary intermediate lobe of Xenopus laevis was explored. Mass spectra were obtained with such a high sensitivity of detection that the peptides could be identified in a single melanotrope cell. In addition to known POMC processing products of the Xenopus melanotrope cell, the presence of previously unidentified POMC-derived peptides was demonstrated. Together these POMC processing products accounted for the entire length of the POMC precursor. Furthermore, Xenopus possesses two genes for POMC and the sensitivity and accuracy of the MALDI-MS technique allowed identification of processing products of both the POMCA and POMCB gene. In addition, differences were obtained between the mass spectra of melanotrope cells from Xenopus laevis adapted to different conditions of background illumination. These results show that MALDI-MS is a valuable tool in the study of the expression of peptides in single (neuroendocrine) cells.
van Strien
F J
FJ
Department of Animal Physiology, Nijmengen Institute for Neurosciences, Subfaculty of Biology, University of Nijmegen, The Netherlands.
Jespersen
S
S
van der Greef
J
J
Jenks
B G
BG
Roubos
E W
EW
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D009074:Melanocyte-Stimulating Hormones / Q000096:biosynthesis / Q000737:chemistry; D008544:Melanocytes; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014982:Xenopus laevis
D000596:Amino Acids; D010446:Peptide Fragments; D011333:Pro-Opiomelanocortin; D009074:Melanocyte-Stimulating Hormones