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• W1986036446 abstract "Abstract Synaptic transmission depends on the efficient loading of transmitters into synaptic vesicles by vesicular neurotransmitter transporters. The vesicular monoamine transporter‐2 (VMAT2) is essential for loading monoamines into vesicles and maintaining normal neurotransmission. In an effort to understand the regulatory mechanisms associated with VMAT2, we have embarked upon a systematic search for interacting proteins. Glutathione‐S‐transferase pull‐down assays combined with mass spectrometry led to the identification of the 70‐kDa heat shock cognate protein (Hsc70) as a VMAT2 interacting protein. Co‐immunoprecipitation experiments in brain tissue and heterologous cells confirmed this interaction. A direct binding was observed between the amino terminus and the third cytoplasmic loop of VMAT2, as well as, a region containing the substrate binding and the carboxy‐terminal domains of Hsc70. Furthermore, VMAT2 and Hsc70 co‐fractionated with purified synaptic vesicles obtained from a sucrose gradient, suggesting that this interaction occurs at the synaptic vesicle membrane. The functional significance of this novel VMAT2/Hsc70 interaction was examined by performing vesicular uptake assays in heterologous cells and purified synaptic vesicles from brain tissue. Recombinant Hsc70 produced a dose‐dependent inhibition of VMAT2 activity. This effect was mimicked by the closely related Hsp70 protein. In contrast, VMAT2 activity was not altered in the presence of previously denatured Hsc70 or Hsp70, as well as the unrelated Hsp60 protein; confirming the specificity of the Hsc70 effect. Finally, a purified Hsc70 fragment that binds VMAT2 was sufficient to inhibit VMAT2 activity in synaptic vesicles. Our results suggest an important role for Hsc70 in VMAT2 function and regulation." @default.
• W1986036446 created "2016-06-24" @default.
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• W1986036446 date "2009-06-25" @default.
• W1986036446 modified "2023-10-17" @default.
• W1986036446 title "The molecular chaperone Hsc70 interacts with the vesicular monoamine transporter-2" @default.
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• W1986036446 doi "https://doi.org/10.1111/j.1471-4159.2009.06135.x" @default.
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