Abstract:We have investigated whether surface plasmon resonance (SPR) sensors based on the wavelength interrogation are able to analyze protein interactions on protein arrays. The spectral SPR sensor was self-constructed and its detection limit, expressed as the minimal refractive index variation, was calculated to be 6.6x10(-5) with the signal fluctuation of 1.0x10(-5). The protein array surface was modified by a mixed thiol monolayer to immobilize proteins. Protein arrays were analyzed by the line-scanning mode of th… Show more
“…Expression screenings in co-culture systems represent a useful method for the identification of novel families of synaptogenic molecules (Paradis et al, 2007; Linhoff et al, 2009). On a larger scale, existing proteomic methods, such as mass spectrometry and protein array tools, can be applied to reveal protein–protein interactions (Husi et al, 2000; Schweitzer et al, 2003; Yuk et al, 2004; Collins and Choudhary, 2008). Bioinformatics tools could also be employed to generate hypothesis of interactions that could be verified experimentally and to construct interaction maps and models that could be used to predict the effects of mutations of single proteins (Armstrong et al, 2006).…”
Section: A Molecular Catalog Of Inhibitory Synapsesmentioning
GABAergic synapses exhibit a high degree of subcellular and molecular specialization, which contrasts with their apparent simplicity in ultrastructural appearance. Indeed, when observed in the electron microscope, GABAergic synapses fit in the symmetric, or Gray’s type II category, being characterized by a relatively simple postsynaptic specialization. The inhibitory postsynaptic density cannot be readily isolated, and progress in understanding its molecular composition has lagged behind that of excitatory synapses. However, recent studies have brought significant progress in the identification of new synaptic proteins, revealing an unexpected complexity in the molecular machinery that regulates GABAergic synaptogenesis. In this article, we provide an overview of the molecular diversity of GABAergic synapses, and we consider how synapse specificity may be encoded by selective trans-synaptic interactions between pre- and postsynaptic adhesion molecules and secreted factors that reside in the synaptic cleft. We also discuss the importance of developing cataloguing tools that could be used to decipher the molecular diversity of synapses and to predict alterations of inhibitory transmission in the course of neurological diseases.
“…Expression screenings in co-culture systems represent a useful method for the identification of novel families of synaptogenic molecules (Paradis et al, 2007; Linhoff et al, 2009). On a larger scale, existing proteomic methods, such as mass spectrometry and protein array tools, can be applied to reveal protein–protein interactions (Husi et al, 2000; Schweitzer et al, 2003; Yuk et al, 2004; Collins and Choudhary, 2008). Bioinformatics tools could also be employed to generate hypothesis of interactions that could be verified experimentally and to construct interaction maps and models that could be used to predict the effects of mutations of single proteins (Armstrong et al, 2006).…”
Section: A Molecular Catalog Of Inhibitory Synapsesmentioning
GABAergic synapses exhibit a high degree of subcellular and molecular specialization, which contrasts with their apparent simplicity in ultrastructural appearance. Indeed, when observed in the electron microscope, GABAergic synapses fit in the symmetric, or Gray’s type II category, being characterized by a relatively simple postsynaptic specialization. The inhibitory postsynaptic density cannot be readily isolated, and progress in understanding its molecular composition has lagged behind that of excitatory synapses. However, recent studies have brought significant progress in the identification of new synaptic proteins, revealing an unexpected complexity in the molecular machinery that regulates GABAergic synaptogenesis. In this article, we provide an overview of the molecular diversity of GABAergic synapses, and we consider how synapse specificity may be encoded by selective trans-synaptic interactions between pre- and postsynaptic adhesion molecules and secreted factors that reside in the synaptic cleft. We also discuss the importance of developing cataloguing tools that could be used to decipher the molecular diversity of synapses and to predict alterations of inhibitory transmission in the course of neurological diseases.
“…Gold arrays which have 50 spots (diameter of each spot: 2 mm) were fabricated and surfaces modified according to a previous report [24]. To fabricate a self-assembled monolayer, Au surfaces were modified with a mixed thiol solution of 1 mM 11-mercaptoundecanoic acid and 2 mM mercaptohexanol in ethanol for 16 h. For amine coupling between the carboxy group of 11-mercaptoundecanoic acid and the amine groups of proteins, the carboxy group of 11-mercaptoundecanoic acid was activated by treating the thiol-modified gold surfaces with a solution of 50 mM Nhydroxysuccinimide and 200 mM N-ethyl-N 0 -(dimethylaminopropyl)-carbodiimide for 10 min.…”
“…Recently, with the development of high-throughput techniques (such as microarrays) [12], angular SPR imaging sensors with multiple channels are developed based on twodimensional photoelectric conversion devices (such as CCD and CMOS cameras) [8][9][10]13]. Since the angular SPR sensors require monochromatic light sources, lasers are often chosen because of their excellent monochromaticity, nice parallelism, and high intensity.…”
The speckle effects caused by the coherence of the laser light sources have restricted the performances of angular surface plasmon resonance (SPR) imaging sensors, which are widely used due to their wide linear rage, high refractive index (RI) resolution, and simple structure. In this work, we experimentally studied the influence of two common despeckle methods (laser with a rotating diffuser and an incoherent light-emitting diodes (LED) light source) on the RI resolution of an angular SPR imaging sensor. We found that the LED light source with a band-pass filter could not only eliminate the speckle effect completely but also provide better RI resolution than the other despeckle method. This low-cost specklefree method has the potential to promote the development and the industrialization of angular SPR imaging sensors.
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