van Strien F J, Jespersen S, van der Greef J, Jenks B G, Roubos E W
Department of Animal Physiology, Nijmengen Institute for Neurosciences, Subfaculty of Biology, University of Nijmegen, The Netherlands.
FEBS Lett. 1996 Jan 29;379(2):165-70. doi: 10.1016/0014-5793(95)01503-5.
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.
研究了基质辅助激光解吸/电离质谱（MALDI-MS）在鉴定非洲爪蟾垂体中间叶黑素细胞中阿片促黑素皮质素原（POMC）加工产物方面的应用。获得的质谱具有如此高的检测灵敏度，以至于可以在单个黑素细胞中鉴定出这些肽。除了非洲爪蟾黑素细胞中已知的POMC加工产物外，还证明了存在以前未鉴定的POMC衍生肽。这些POMC加工产物加起来占POMC前体的全长。此外，非洲爪蟾有两个POMC基因，MALDI-MS技术的灵敏度和准确性使得能够鉴定POMCA和POMCB基因的加工产物。此外，适应不同背景光照条件的非洲爪蟾黑素细胞的质谱之间存在差异。这些结果表明，MALDI-MS是研究单个（神经内分泌）细胞中肽表达的有价值工具。
