Synapsins as major neuronal Ca2+/S100A1-interacting proteins
| Autor: | Heierhorst, J, Mitchelhill, K I, Mann, R J, Tiganis, T, Czernik, A J, Greengard, P, Kemp, B E |
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| Jazyk: | angličtina |
| Rok vydání: | 1999 |
| Předmět: |
Brain Chemistry
Neurons Calcium-Binding Proteins Molecular Sequence Data S100 Proteins Synapsins PC12 Cells Chromatography Affinity Cell Compartmentation Rats Adenosine Triphosphate nervous system Sequence Analysis Protein Neurites Animals Protein Isoforms Calcium Amino Acid Sequence Research Article Protein Binding |
| Popis: | The mammalian S100A1 protein can activate the invertebrate myosin-associated giant protein kinase twitchin in a Ca(2+)-dependent manner by more than 1000-fold in vitro; however, no mammalian S100-dependent protein kinases are known. In an attempt to identify novel mammalian Ca(2+)/S100A1-regulated protein kinases, brain extracts were subjected to combined Ca(2+)-dependent affinity chromatography with S100A1 and an ATP analogue. This resulted in the purification to near-homogeneity of the four major synapsin isoforms Ia, Ib, IIa and IIb. All four synapsins were specifically affinity-labelled with the ATP analogue 5'-p-fluorosulphonylbenzoyladenosine. S100A1 bound to immobilized synapsin IIa in BIAcore experiments in a Ca(2+)-dependent and Zn(2+)-enhanced manner with submicromolar affinity; this interaction could be competed for with synthetic peptides of the proposed S100A1-binding sites of synapsin. Double-labelling confocal immunofluorescence microscopy demonstrated that synapsins and S100A1 are both present in the soma and neurites of PC12 cells, indicating their potential to interact in neurons in vivo. |
| Databáze: | OpenAIRE |
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