The effect of calcium levels on synaptic proteins. A study on VAT-1 from Torpedo.
In this study we compare major synaptic proteins from Torpedo electric organ to their homologues from mammalian brain. Most of these proteins are members of small gene families. We demonstrate a high degree of evolutionary conservation of most synaptic proteins. However, in the electric organ each gene family is represented only by a single member. We focus on VAT-1, a major protein of the vesicle membrane in Torpedo. VAT-1 is located on the synaptic vesicle membrane and is highly concentrated on the plasma membrane following the application of alpha-latrotoxin. Taking advantage of the relative simplicity of Torpedo synapses, we performed an in vitro study on the properties of VAT-1 affected by changes in Ca2+ levels. VAT-1 is a low affinity Ca2+ binding protein whose ability to bind Ca2+ resides mainly, but not entirely, on the carboxy-terminal domain of the protein. In the presence of Ca2+, the protein is organized in a high molecular mass complex, which is destabilized by depleting Ca2+. This effect occurs only by chelating Ca2+ ions, but not with other divalent ions. VAT-1 is not complexed to any of the proteins which were implicated in the docking/fusion complex such as VAMP, synaptophysin or syntaxin, regardless of Ca2+ levels. Dependence of the stability of protein complexes on Ca2+ levels is also demonstrated on Torpedo n-Sec1. The possible physiological implications of such Ca2+ dependence are discussed.
Department of Biological Chemistry, Alexander Silberman Institute of Life Sciences, Hebrew University of Jerusalem, Israel.
M
M
Linial
O
O
Levius
N
N
Ilouz
D
D
Parnas
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D004557:Electric Organ / Q000294:innervation / Q000378:metabolism* / Q000648:ultrastructure; D004926:Escherichia coli / Q000378:metabolism; D015151:Immunoblotting; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000502:physiology*; D011233:Precipitin Tests; D013570:Synaptic Membranes / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D014101:Torpedo / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C063277:VAT-1 protein, Torpedo; D002118:Calcium