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  • 1
    Online Resource
    Online Resource
    Berlin, Heidelberg :Springer Berlin / Heidelberg,
    Keywords: Nanotechnology. ; Electronic books.
    Type of Medium: Online Resource
    Pages: 1 online resource (227 pages)
    Edition: 1st ed.
    ISBN: 9783662433676
    Series Statement: Advances in Atom and Single Molecule Machines Series
    DDC: 620.5
    Language: English
    Note: Intro -- Preface -- Contents -- Contributors -- 1 Introduction to Single-Molecule Analysis and Computation: The Focus Project -- References -- 2 Computer-Generated Holographic Beams for the Investigation of the Molecular and Circuit Function -- Abstract -- 1…Introduction -- 2…Background and Computational Aspects in CGH -- 2.1 Computation of the Phase Maps -- 3…Hardware for CGH: LCoS-SLM -- 3.1 LCoS-SLM: Working Principle -- 3.2 LCoS-SLM: Temporal Features and Overdriving Procedures -- 3.3 LCoS-SLM: Design of the Device -- 3.4 LCoS-SLM: Optical Properties -- 4…Optical Configurations for CGH -- 4.1 Basic Configuration for CGH -- 4.2 Integration of CGH into Imaging Systems -- 4.3 Optical Figures of CGH -- 4.4 Peculiar Aspects of CGH Beams for Bidimensional Extended Patterns -- 5…Applications and Perspective for CGH -- References -- 3 Super-Resolution Fluorescence Optical Microscopy: Targeted and Stochastic Read-Out Approaches -- Abstract -- 1…Introduction -- 2…Toward Super-Resolution Imaging of Thick ( greaterthan 50 Mu m) Biological Specimens -- 3…Single-Molecule Localization Techniques (Palm-Like) -- 4…Stimulated Emission Depletion (Sted-Like) -- 5…Perspective Note -- References -- 4 Superhydrophobic Devices Molecular Detection -- Abstract -- 1…Introduction -- 1.1 Single-Molecule Detection -- 1.2 Superhydrophobic Surfaces -- 2…Fabrication -- 2.1 Superhydrophobic Pillared Substrates -- 2.2 Polymeric Superhydrophobic Substrates -- 3…Applications -- 3.1 Molecular Detection of Organic and Inorganic Sample's Structural Information Through In-situ X-ray Diffraction -- 3.2 Superhydrophobic Substrate for Raman Detection -- 3.3 Superhydrophobic Substrates for Background-Free TEM DNA Characterization and for Nucleic Acid Pattern Generation -- References -- 5 Tip-Assisted Optical Nanoscopy for Single-Molecule Activation and Detection -- Abstract. , 1…An Overview -- 1.1 A Brief History of the Tip-Assisted Technique -- 2…Theoretical Background -- 2.1 The Evanescent Field -- 2.2 Localized Surface Plasmon Tip -- 2.3 Plasmonic Antenna Tip -- 3…Experimental Setups and Methods -- 3.1 Tips and Tips Engineering -- 3.2 Microscopes Design -- 4…Applications of TERS -- References -- 6 DNA as Nanostructuring Element for Design of Functional Devices -- Abstract -- 1…Introduction -- 2…DNA-Directed Immobilization: Use of Chemically Synthesized Short DNA -- 2.1 DNA as an Immobilization Tool -- 2.2 DNA--Nanoparticle Conjugates -- 3…DNA Origami: Combination of Natural and Chemically Synthesized DNA -- 4…DNA in Design of Thin Films and Optoelectronic Devices: Use of DNA from Natural Sources -- 5…DNA-Based Materials -- 5.1 DNA-Based Hydrogels -- 5.2 DNA-Polymeric Hybrid Materials -- 5.3 DNA Side-Chain Polymers -- 6…Conclusion -- References -- 7 Fast Force Clamp in Optical Tweezers: A Tool to Study the Kinetics of Molecular Reactions -- Abstract -- 1…Single-Molecule Mechanics with Optical Tweezers -- 2…The Dual-Laser Optical Tweezers -- 2.1 Mounting the Specimen in the Experimental Chamber Between the Force and Length Transducers -- 2.2 Calibration of Force -- 2.3 Mechanical Protocols: Length Versus Force Clamp -- 3…Transition Kinetics of the DNA Elongation Revealed by Force Steps -- 4…The Structure of the DNA Overstretching Transition -- 5…Assessing the Force Clamp Description of DNA Overstretching Kinetics: Comparison Between Non-equilibrium and Equilibrium Results -- A.x(118). Appendix: Influence of Viscosity on the Kinetics of the Elongation--Untwisting of the ds-DNA During the Overstretching Transition -- A.x(118).0 Drag Produced on the Trapped Bead by the Viscosity of the Medium -- A.x(118).0 Rotational Drag of the Molecule While Untwisting -- References. , 8 Investigating Adhesion Proteins by Single Cell Force Spectroscopy -- Abstract -- 1…Introduction -- 2…Force-Spectroscopy Measurements and Experimental Details -- 3…Analysis of Force-Distance Curve -- 4…The Advantages of SCFS in the Study of Single Adhesion Proteins -- 5…Quantifying Cell Adhesion at Single Molecule Scale -- 6…Future Perspectives -- References -- 9 Photoswitchable Ion Channels and Receptors -- Abstract -- 1…Introduction -- 2…Pentameric Ligand-Gated Receptors -- 2.1 Introduction -- 2.2 Nicotinic Acetylcholine Receptors (nAChR) -- 2.3 Ionotropic gamma -Aminobutyric Acid Receptor (GABAA) -- 3…Tetrameric Ligand-Gated Receptors -- 3.1 Introduction -- 3.2 Kainate Receptors -- 3.3 AMPA Receptors -- 3.4 Bacterial iGluRs -- 3.5 Metabotropic Glutamate Receptors -- 3.6 Applications -- 4…Trimeric Ligand-Gated Receptors -- 5…Potassium Channels -- 5.1 Introduction -- 5.2 Kv Family -- 5.2.1 2P-Potassium Channels -- 6…Transient Receptor Potential Channels -- 7…Future Challenges -- References -- 10 The Use of Light-Sensitive Organic Semiconductors to Manipulate Neuronal Activity -- Abstract -- 1…Introduction -- 2…Optical Techniques Modulating Neuronal Activity -- 3…Organic Semiconductive Polymers -- 4…Modulation of Membrane Potential Using Light-Sensitive Polymers -- 5…Application of Organic Photovoltaic Devices in Retinal Degenerative Diseases -- 6…Future Research Direction -- 7…Conclusions -- References -- 11 Probing the Lateral Diffusion of Individual Neurotransmitter Receptors -- Abstract -- 1…Introduction -- 2…Measuring the Lateral Diffusion of Neurotransmitter Receptors -- 2.1 Population Diffusion Measurements -- 2.2 Single-particle Tracking Techniques -- 3…Role of Receptor Diffusion in Long-Term Synaptic Plasticity -- 4…Implications of Receptor Diffusion in Fast Synaptic Signaling -- 5…Future Perspectives -- References.
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  • 2
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Focal adhesion kinase (FAK) and the related proline-rich tyrosine kinase 2 (PYK2) are non-receptor protein tyrosine kinases that transduce extracellular signals through the activation of Src family kinases and are highly enriched in neurones. To further elucidate the regulation of FAK and PYK2 in nervous tissue, we investigated their distribution in brain subcellular fractions and analysed their translocation between membrane and cytosolic compartments. We have found that FAK and PYK2 are present in a small membrane-associated pool and a larger cytosolic pool in various neuronal compartments including nerve terminals. In intact nerve terminals, inhibition of Src kinases inhibited the membrane association of FAK, but not of PYK2, whereas tyrosine phosphatase inhibition sharply increased the membrane association of both FAK and PYK2. Disruption of the actin cytoskeleton was followed by a decrease in the membrane-associated pool of FAK, but not of PYK2. For both kinases, a significant correlation was found between autophosphorylation and membrane association. The data indicate that FAK and PYK2 are present in nerve terminals and that the membrane association of FAK is regulated by both phosphorylation and actin assembly, whereas that of PKY2 is primarily dependent on its phosphorylation state.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: VAMP/synaptobrevin (SYB), an integral membrane protein of small synaptic vesicles, is specifically cleaved by tetanus neurotoxin and botulinum neurotoxins B, D, F, and G and is thought to play an important role in the docking and/or fusion of synaptic vesicles with the presynaptic membrane. Potential phosphorylation sites for various kinases are present in SYB sequence. We have studied whether SYB is a substrate for protein kinases that are present in nerve terminals and known to modulate neurotransmitter release. SYB can be phosphorylated within the same vesicle by endogenous Ca2+/calmodulin-dependent protein kinase II (CaMKII) associated with synaptic vesicles. This phosphorylation reaction occurs rapidly and involves serine and threonine residues in the cytoplasmic region of SYB. Similarly to CaMKII, a casein kinase II (CasKII) activity copurifying with synaptic vesicles is able to phosphorylate SYB selectively on serine residues of the cytoplasmic region. This phosphorylation reaction is markedly stimulated by sphingosine, a sphingolipid known to activate CasKII and to inhibit CaMKII and protein kinase C. The results show that SYB is a potential substrate for protein kinases involved in the regulation of neurotransmitter release and open the possibility that phosphorylation of SYB plays a role in modulating the molecular interactions between synaptic vesicles and the presynaptic membrane.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 57 (1991), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Generation of antibodies and direct protein sequencing were used to identify and characterize proteins associated with highly purified synaptic vesicles from rat brain. A protein doublet of low abundance of 119 and 124 kDa apparent molecular mass [synaptic vesicle-associated phosphoprotein with a molecular mass of 120 kDa (SVAPP-120)] was identified using polyclonal antibodies. SVAPP-120 was found to copurify with synaptic vesicles and to be enriched in the purified synaptic vesicle fraction to the same extent as synapsin I. Like synapsin I, SVAPP-120 is not an integral membrane protein because it was released from synaptic vesicles by high salt concentrations. This protein was demonstrated to be brain specific, and its distribution in various brain regions paralleled the distribution of synapsin I and synaptophysin. During the postnatal development of the rat cortex and cerebellum, its expression correlated with synaptogenesis. SVAPP-120 was demonstrated to be a phosphoprotein both in vivo and in vitro. It was shown to be phosphorylated on serine and to a lesser extent on threonine residues. These results provide evidence that SVAPP-120 represents a novel synaptic vesicle-associated phosphoprotein. In addition, aldolase, a glycolytic enzyme, and αc-adaptin, a clathrin assembly-promoting protein, were identified on purified synaptic vesicles by direct protein sequencing.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The phosphorylation of specific proteins in pineal homogenate was analyzed by sodium dodecyl sulfate-poly-acrylamide gel electrophoresis and autoradiography. Cyclic AMP had the capacity to stimulate in a dose-dependent manner the incorporation of 32P in protein bands of apparent molecular weights 59K, 56K, and 35K with a maximal effect at 1 μM. On the other hand, calcium alone did not induce a marked increase in 32P incorporation with the exception of a dose-dependent phosphorylation of a 46K protein with a peak effect at 0.2 mM calcium concentration. The addition of exogenous calmodulin enhanced 32P incorporation in proteins migrating in the 62K and 52K regions, an effect that was antagonized by the calmodulin inhibitor trifluoperazine. However, also under these conditions, the stimulation of pineal protein phosphorylation was rather weak compared to that observed in other brain areas. In an attempt to investigate the functional changes of these biochemical processes during environmental lighting and ad-renergic stimulation, it was found that the administration of (-)-isoproterenol (5 mg/kg, s.c), a β-receptor agonist, induced a clear-cut enhancement of 32P incorporation into the cyclic AMP-sensitive 59K and 56K proteins only in animals exposed for 18 h to the light, whereas it was almost ineffective in those kept in the dark for the same period. This effect was antagonized by (—)-propranolol pretreatment (20 mg/ kg), suggesting that the changes in cyclic AMP-dependent protein phosphorylation observed in supersensitive pineals may represent a β-receptor mediated process. Thus the cyclic AMP-dependent protein kinase activity seems to be correlated with the sensitivity state of the β-adrenergic receptor system and may take part in the development of pineal su-persensitivity. On the other hand, the phosphorylation of the 46K calcium-sensitive protein was completely unaffected, confirming the action specificity of the adrenergic system on cyclic AMP-induced pineal protein phosphorylation.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Several cytokines have short-term effects on synaptic transmission and plasticity that are thought to be mediated by the activation of intracellular protein kinases. We have studied the effects of interleukin-6 (IL-6) on the expression of paired pulse facilitation (PPF), posttetanic potentiation (PTP), and long-term potentiation (LTP) in the CA1 region of the hippocampus as well as on the activation of the signal transducer and activator of transcription-3 (STAT3), the mitogen-activated protein kinase ERK (MAPK/ERK), and the stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK). IL-6 induced a marked and dose-dependent decrease in the expression of PTP and LTP that could be counteracted by the simultaneous treatment with the tyrosine kinase inhibitor lavendustin A (LavA) but did not significantly affect PPF. The IL-6-induced inhibition of PTP and LTP was accompanied by a simulation of STAT3 tyrosine phosphorylation and an inhibition of MAPK/ERK dual phosphorylation, in the absence of changes in the state of activation of SAPK/JNK. Both effects of IL-6 on STAT3 and MAPK/ERK activation were effectively counteracted by LavA treatment. The results indicate the tyrosine kinases and MAPK/ERK are involved in hippocampal synaptic plasticity and may represent preferential intracellular targets for the actions of IL-6 in the adult nervous system.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Immunophilins are ubiquitous enzymes responsible for proline isomerisation during protein synthesis and for the chaperoning of several membrane proteins. These activities can be blocked by the immunosuppressants cyclosporin A, FK506 and rapamycin. It has been shown that all three immunosuppressants have neurotrophic activity and can modulate neurotransmitter release, but the molecular basis of these effects is currently unknown. Here, we show that synapsin I, a synaptic vesicle-associated protein, can be purified from Torpedo cholinergic synaptosomes through its affinity to cyclophilin B, an immunophilin that is particularly abundant in brain. The interaction is direct and conserved in mammals, and shows a dissociation constant of about 0.5 µmin vitro. The binding between the two proteins can be disrupted by cyclosporin A and inhibited by physiological concentrations of ATP. Furthermore, cyclophilin B co-localizes with synapsin I in rat synaptic vesicle fractions and its levels in synaptic vesicle-containing fractions are decreased in synapsin knockout mice. These results suggest that immunophilins are involved in the complex protein networks operating at the presynaptic level and implicate the interaction between cyclophilin B and synapsins in presynaptic function.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The Ca2+-sensor protein S100A1 was recently shown to bind in vitro to synapsins, a family of synaptic vesicle phosphoproteins involved in the regulation of neurotransmitter release. In this paper, we analyzed the distribution of S100A1 and synapsin I in the CNS and investigated the effects of the S100A1/synapsin binding on the synapsin functional properties. Subcellular fractionation of rat brain homogenate revealed that S100A1 is present in the soluble fraction of isolated nerve endings. Confocal laser scanning microscopy and immunogold immunocytochemistry demonstrated that S100A1 and synapsin codistribute in a subpopulation (5–20%) of nerve terminals in the mouse cerebral and cerebellar cortices. By forming heterocomplexes with either dephosphorylated or phosphorylated synapsin I, S100A1 caused a dose- and Ca2+-dependent inhibition of synapsin-induced F-actin bundling and abolished synapsin dimerization, without affecting the binding of synapsin to F-actin, G-actin or synaptic vesicles. These data indicate that: (i) synapsins and S100A1 can interact in the nerve terminals where they are coexpresssed; (ii) S100A1 is unable to bind to SV-associated synapsin I and may function as a cytoplasmic store of monomeric synapsin I; and (iii) synapsin dimerization and interaction with S100A1 are mutually exclusive, suggesting an involvement of S100A1 in the Ca2+-dependent regulation of synaptic vesicle trafficking.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 31 (1992), S. 4268-4275 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The synapsins are a family of synaptic vesicle phosphoproteins which play a key role in the regulation of neurotransmitter release and synapse formation. In the case of synapsin I, these biological properties have been attributed to its ability to interact with both synaptic vesicles and the actin-based cytoskeleton. Although synapsin II shares some of the biological properties of synapsin I, much less is known of its molecular properties. We have investigated the interactions of recombinant rat synapsin Ila with monomeric and filamentous actin and the sensitivity of those interactions to phosphorylation, and found that: i) dephosphotylated synapsin II stimulates actin polymerization by binding to actin monomers and forming actively elongating nuclei and by facilitating the spontaneous nucleation/elongation processes; ii) dephosphorylated synapsin II induces the formation of thick and ordered bundles of actin filaments with greater potency than synapsin I; iii) phosphorylation by protein kinase A markedly inhibits the ability of synapsin II to interact with both actin monomers and filaments. The results indicate that the interactions of synapsin II with actin are similar but not identical to those of synapsin I and suggest that synapsin II may play a major structural role in mature and developing nerve terminals, which is only partially overlapping with the role played by synapsin I.
    Type of Medium: Electronic Resource
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