11 research outputs found

    SNARE protein recycling by ?SNAP and ?SNAP supports synaptic vesicle priming

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    Neurotransmitter release proceeds by Ca(2+)-triggered, SNARE-complex-dependent synaptic vesicle fusion. After fusion, the ATPase NSF and its cofactors ?- and ?SNAP disassemble SNARE complexes, thereby recycling individual SNAREs for subsequent fusion reactions. We examined the effects of genetic perturbation of ?- and ?SNAP expression on synaptic vesicle exocytosis, employing a new Ca(2+) uncaging protocol to study synaptic vesicle trafficking, priming, and fusion in small glutamatergic synapses of hippocampal neurons. By characterizing this protocol, we show that synchronous and asynchronous transmitter release involve different Ca(2+) sensors and are not caused by distinct releasable vesicle pools, and that tonic transmitter release is due to ongoing priming and fusion of new synaptic vesicles during high synaptic activity. Our analysis of ?- and ?SNAP deletion mutant neurons shows that the two NSF cofactors support synaptic vesicle priming by determining the availability of free SNARE components, particularly during phases of high synaptic activit

    Elucidating the early steps in photoinitiated radical polymerization via femtosecond pump-probe experiments and DFT calculations

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    The excited states and dynamics of the three triplet radical photoinitiators benzoin (2-hydroxy-1,2-diphenylethanone, Bz), 2,4,6-trimethylbenzoin (2-hydroxy-1-mesityl-2-phenylethanone, TMB), and mesitil (1,2-bis(2,4,6-trimethylphenyl)-1,2-ethanedione, Me)-employed in our previous studies for quantifying net initiation efficiencies in pulsed laser polymerization with methacrylate monomers [Voll, D.; Junkers, T.; Barner-Kowollik, C. Macromolecules2011, 44, 2542-2551]-are investigated via both femtosecond transient absorption (TA) spectroscopy and density functional theory (DFT) methods to elucidate the underlying mechanisms causing different initiation efficiencies when excited at 351 nm. Bz and TMB are found to have very similar properties in the calculated excited states as well as in the experimentally observed dynamics. After excitation into the first excited singlet state (S 1) Bz and TMB undergo rapid intersystem crossing (ISC). The ISC can compete with ultrafast internal conversion (IC) processes because an excited triplet state (T n) of nearly the same energy is present in both cases. ISC is therefore the dominating depopulation channel of S 1, and subsequent α-cleavage to produce radicals takes place on the picosecond time scale. In contrast, Me is excited into the second excited singlet state (S 2). In this case no isoenergetic triplet state is available, which inhibits ISC from competing with ultrafast deactivation processes. ISC is therefore assigned to be a minor deactivation channel in Me. Employing these findings, quantitative photoinitiation efficiency relations of Bz, TMB, and Me obtained by pulsed laser polymerization can be directly correlated with the relative TA intensities found in the femtosecond experiments. The ISC efficiency is thus a critical parameter for evaluating the overall photoinitiation efficiency and demonstrates that the employment of the herein presented method represents a powerful tool for attaining a quantitative picture on the suitability of a photoinitiator. © 2012 American Chemical Society

    Functionality of muscle proteins in gelation mechanisms of structured meat products

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