The mammalian S100A1 protein can activate the invertebrate myosin-associated giant protein kinase twitchin in a Ca2+-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 Ca2+/S100A1-regulated protein kinases, brain extracts were subjected to combined Ca2+-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 Ca2+-dependent and Zn2+-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.
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December 1999
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Research Article|
November 24 1999
Synapsins as major neuronal Ca2+/S100A1-interacting proteins
Jörg HEIERHORST;
Jörg HEIERHORST
1
*St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, VIC 3065, Australia
1To whom correspondence should be addressed (e-mail heier@ariel.its.unimelb.edu.au).
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Ken I. MITCHELHILL;
Ken I. MITCHELHILL
*St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, VIC 3065, Australia
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Richard J. MANN;
Richard J. MANN
*St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, VIC 3065, Australia
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Tony TIGANIS;
Tony TIGANIS
*St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, VIC 3065, Australia
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Andrew J. CZERNIK;
Andrew J. CZERNIK
†Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, 1230 York Avenue, New York, NY 10021, U.S.A.
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Paul GREENGARD;
Paul GREENGARD
†Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, 1230 York Avenue, New York, NY 10021, U.S.A.
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Bruce E. KEMP
Bruce E. KEMP
*St. Vincent's Institute of Medical Research, 41 Victoria Parade, Fitzroy, VIC 3065, Australia
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Publisher: Portland Press Ltd
Received:
May 28 1999
Revision Received:
August 09 1999
Accepted:
September 10 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 344 (2): 577–583.
Article history
Received:
May 28 1999
Revision Received:
August 09 1999
Accepted:
September 10 1999
Citation
Jörg HEIERHORST, Ken I. MITCHELHILL, Richard J. MANN, Tony TIGANIS, Andrew J. CZERNIK, Paul GREENGARD, Bruce E. KEMP; Synapsins as major neuronal Ca2+/S100A1-interacting proteins. Biochem J 1 December 1999; 344 (2): 577–583. doi: https://doi.org/10.1042/bj3440577
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