Voltage-dependent N-type Ca2+ channel activity regulates the interaction between FGF-1 and S100A13 for stress-induced non-vesicular release.
1. The Ca(2+)-mediated regulation of interaction between FGF-1 and S100A13 in NG108-15 cells was studied. When the stress by depriving B27 supplement from the culture was given, cellular levels of both proteins were decreased, while their releases were significantly increased within 3 h. These stress-induced changes were all abolished by amlexanox, an anti-allergic drug. 2. These releases were significantly inhibited by the addition of EGTA or BAPTA-AM, cellular or extracellular Ca(2+)-chelating agent, respectively. The addition of omega-conotoxin GVIA, a N-type Ca(2+)-channel blocker caused a complete inhibition of the release, while increased the cytosolic levels of both proteins. However, omega-conotoxin MVIIC, the non-N-type Ca(2+)-channel blocker was ineffective. 3. In NG108-15 cells, which had been transfected with Venus-FGF-1 and CFP-S100A13, the supplement-deprivation stress caused several spike-type fluorescence resonance energy transfer (FRET) signals, suggesting that both proteins showing interaction would be immediately released. These spikes were completely abolished by the addition of omega-conotoxin GVIA. However, the addition of amlexanox caused bell-shaped FRET signals without spikes. 4. Thus, it is suggested that the interaction between FGF-1 and S100A13 responsible for stress-induced non-vesicular release is dependent of Ca(2+)-influx through N-type Ca(2+)-channels.
Matsunaga
Hayato
H
Division of Molecular Pharmacology and Neuroscience, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. ueda@net.nagasaki-u.ac.jp
Ueda
Hiroshi
H
D000631:Aminopyridines / Q000378:metabolism; D000818:Animals; D018926:Anti-Allergic Agents / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000378:metabolism; D020864:Calcium Channels, N-Type / Q000378:metabolism*; D002460:Cell Line; D002614:Chelating Agents / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D051381:Rats; D009418:S100 Proteins / Q000235:genetics / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*; D020866:omega-Conotoxin GVIA / Q000378:metabolism
D000631:Aminopyridines; D018926:Anti-Allergic Agents; D002121:Calcium Channel Blockers; D020864:Calcium Channels, N-Type; D002614:Chelating Agents; D008164:Luminescent Proteins; D009418:S100 Proteins; D016220:Fibroblast Growth Factor 1; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D020866:omega-Conotoxin GVIA; C045742:amlexanox; D002118:Calcium