Direct mass spectrometric peptide profiling and sequencing of single neurons reveals differential peptide patterns in a small neuronal network
| Autor: | Sabine Spijker, J. Van Minnen, Alma L. Burlingame, Connie R. Jimenez, K. Dreisewerd, August B. Smit, Ka Wan Li, W. P. M. Geraerts, R. H. Bateman, Robert E. Kingston |
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| Přispěvatelé: | Molecular and Cellular Neurobiology |
| Rok vydání: | 1998 |
| Předmět: |
DNA
Complementary Invertebrate Hormones Molecular Sequence Data Peptide Biology Mass spectrometry Biochemistry law.invention Peptide mass fingerprinting SDG 3 - Good Health and Well-being law Complementary DNA medicine Biological neural network Animals Amino Acid Sequence RNA Messenger Cloning Molecular Protein Precursors Polymerase chain reaction In Situ Hybridization Lymnaea chemistry.chemical_classification Neurons Base Sequence cDNA library Myocardium Neuropeptides Combinatorial chemistry Myocardial Contraction medicine.anatomical_structure chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Neuron |
| Zdroj: | Jimenez, C R, Li, K W, Dreisewerd, K, Spijker, S, Kingston, R, Bateman, R H, Burlingame, A L, Smit, A B, van Minnen, J & Geraerts, W P M 1998, ' Direct mass spectrometric peptide profiling and sequencing of single neurons reveals differential peptide patterns in a small neuronal network ', Biochemistry, vol. 37, pp. 2070-2076 . https://doi.org/10.1021/bi971848b Biochemistry, 37, 2070-2076. American Chemical Society |
| ISSN: | 0006-2960 |
| Popis: | Mass spectrometry (MS) was employed to detect and structurally characterize peptides in two functionally related neurons, named VD1 and RPD2, which form a network involved in the modulation of heartbeat in Lymnaea. Matrix-assisted laser desorption/ionization MS, directly applied to single neurons VD1 and RPD2, showed overlapping yet distinct mass profiles, with a subset of putative peptides specifically present in neuron VD1. Direct tandem MS of a single VD1 neuron revealed the primary structures of the VD1- specific peptides, which were identified as members of the family of small cardioactive peptides. Based on the tandem MS data, a degenerate oligonucleotide was made for use in a polymerase chain reaction strategy to isolate the cDNA encoding the precursor to the small cardioactive peptides from a brain-specific cDNA library. The calculated masses of the mature, posttranslationally modified peptides, as predicted from the corresponding cDNA, agreed with the measured masses of the actual peptides, as detected in single-cell MS analysis. In situ hybridization studies showed that the transcript encoding the precursor is present in VD1, but not in RPD2, thus corroborating the single-cell MS analysis. Finally, the small cardioactive peptides were shown to enhance the contractions of the auricle in vitro. |
| Databáze: | OpenAIRE |
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