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Molecular & Cellular Proteomics 3:558-564, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Identification of Neuropeptides from the Sinus Gland of the Crayfish Orconectes limosus Using Nanoscale On-line Liquid Chromatography Tandem Mass Spectrometry^*

Patrick Bulau^,, Iris Meisen, Tina Schmitz, Rainer Keller and Jasna Peter-Katalini^,¶

From the Institute for Zoophysiology, University of Bonn, Endenicher Allee 11-13, D-53115 Bonn, Germany; and Institute for Medical Physics and Biophysics, Biomedical Analytics, University of Münster, Robert-Koch-Straße 31, D-48149 Münster, Germany

In this article, a novel and sensitive analytical strategy for direct characterization of neuropeptides from the X-Organ-sinus gland neurosecretory system of the crayfish Orconectes limosus is presented. A desalted extract corresponding to 0.5 sinus gland equivalents was analyzed in a nanoflow liquid chromatography system coupled to quadrupole time-of-flight tandem mass spectrometry (nanoLC-QTOF MS/MS). The existence and structural identity of four crustacean hyperglycemic hormone precursor-related peptide variants and two new genetic variants of the pigment-dispersing hormone, not detected by conventional chromatographic systems, molecular cloning, or immunochemical methods before, was revealed. The here-presented approach of the combined LC-QTOF MS/MS technique is a powerful tool to discover new peptide hormones in biological systems, due to its sensitivity, accuracy, and speed.

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