Towards the Realization of Nanobiosensors Based on G‐protein‐coupled Receptors

Pennetta, C.; Акимов, В. Н.; Alfinito, Eleonora; Reggiani, L.; Gorojankina, Tatiana; Minic, Jasmina; Pajot‐Augy, Edith; Persuy, Marie‐Annick; Salesse, Roland; Casuso, Ignacio; Errachid, Abdelhamid; Gomila, Gabriel; Ruiz, Oscar N.; Samitier, J.; Hou, Yanxia; Jaffrezic, N.; Ferrari, Giorgio; Fumagalli, Laura; Sampietro, Marco

doi:10.1002/9783527610419.ntls0041

Cited by 12 publications

(27 citation statements)

References 85 publications

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“…In some recent works [15,16,17], by assigning to each link an impedance, we proposed a new strategy for the investigation of the protein electrical response. Furthermore, on the same ground, also the role of thermal fluctuations was tested [15,16,17].…”

Section: Modelmentioning

confidence: 99%

“…Furthermore, on the same ground, also the role of thermal fluctuations was tested [15,16,17]. Here we use the same strategy by assuming that the amino acids interact electrically among them: The electrical charge transfers between neighboring residues [18], which can also change their electronic polarization [19].…”

Section: Modelmentioning

confidence: 99%

“…More precisely, in this model each link between the nodes i and j corresponds to an elementary RC parallel impedance Z i,j , with R an Ohmic resistor and and C a planar homogeneous capacitor. By assuming that the cross-sectional area of resistor and capacitor is equal to the cross-section area of overlapping spheres, the elementary impedance becomes [16] …”

Section: Modelmentioning

confidence: 99%

“…The relative dielectric constant of the couple of i, j amino acids, ǫ i,j , is expressed in terms of the intrinsic polarizability of the i, j amino acids, α i , α j [16,19] as:…”

Section: Modelmentioning

confidence: 99%

“…Figure 1 reports the difference in the number of links between couples of structures of rhodopsin, native (left) and activated (right), considered here as a function of the interaction radius. The native (Rho) and activated (MetaII) configurations refer to engineered structures obtained from the incomplete 1LN6, 1JFP and 1F88 (chain A) [15,16,17] and contain the same number of amino acids (348). The comparison among the three native representations of rhodopsin, 1U19, 2G87, 2HPY and the engineered one, Rho, shows that: (i) 1U19 and 2G87 exhibit practically the same number of links and, (ii) there are consistent differences between the native and engineered configurations.…”

Section: A the Topological Networkmentioning

confidence: 99%

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A network model to correlate conformational change and the impedance spectrum of single proteins

2008

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One of the main trend in to date research and development is the miniaturization of electronic devices. In this perspective, integrated nanodevices based on proteins or biomolecules are attracting a major interest. In fact, it has been shown that proteins like bacteriorhodopsin and azurin, manifest electrical properties which are promising for the development of active components in the field of molecular electronics. Here we focus on two relevant kinds of proteins: The bovine rhodopsin, prototype of GPCR protein, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer disease. Both these proteins exert their functioning starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different electrical response associated with the different configurations. The model resolution of the electrical response is found able to monitor the structure and the conformational change of the given protein. In this respect, rhodopsin exhibits a better differential response than AChE. This result gives room to different interpretations of the degree of conformational change and in particular supports a recent hypothesis on the existence of a mixed state already in the native configuration of the protein.

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Section: Modelmentioning

confidence: 99%

Section: Modelmentioning

confidence: 99%

Section: Modelmentioning

confidence: 99%

“…The relative dielectric constant of the couple of i, j amino acids, ǫ i,j , is expressed in terms of the intrinsic polarizability of the i, j amino acids, α i , α j [16,19] as:…”

Section: Modelmentioning

confidence: 99%

Section: A the Topological Networkmentioning

confidence: 99%

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A network model to correlate conformational change and the impedance spectrum of single proteins

2008

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Immobilization of Olfactory Receptors Carried by Nanosomes onto a Gold Sensor Surface

Vidić

Hou

2021

Methods in Molecular Biology

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Mammalian olfactory receptors (ORs) constitute the largest family of G-protein coupled receptors, with up to about 1000 different genes per species, each having specific odorant ligands. ORs could be used as sensing elements of highly specific and sensitive bioelectronic hybrid devices such as bioelectronic noses. After optimized immobilization onto the device, natural ORs provide molecular recognition of various odors with their intrinsic sensitivity, discrimination, and detection properties. However, the main difficulties are related to the low expression level of recombinant ORs, their stability and potential loss of activity. Such drawbacks can be successfully overcome in bioelectronic noses integrating nanosomes (nanometric membrane vesicles carrying ORs) that are stably immobilized through a specific antibody. The advantages of such a platform rely on the fact that ORs stay in the natural membrane environment and thus, preserve their full activity. Thanks to their small sizes, nanosomes offer potential for micro-and nano-scale sensor development. In this paper, we summarize the key elements regarding nanosomes production and manipulation and provide an example of their immobilization onto a gold sensor surface. Rat ORI7 is used as representative OR that can be functionally expressed in Saccharomyces cerevisiae. The receptor was not purified but only nanosomes were prepared. Nanosomes were immobilized onto functionalized gold surface using the anti-I7 antibody. Utilization of the antibody provides enrichment of ORI7 on the sensor surface but also uniform and appropriate orientation of the receptors. These features are crucial in optimization of bioelectronic nose' analytical performances.

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Nanobiosensors based on individual olfactory receptors

Akimov

Alfinito

Bausells

et al. 2007

Analog Integr Circ Sig Process

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The animal olfactory system represents the gold standard of biosensors, due to its ability to identify and discriminate thousands of odorant compounds with very low thresholds. Using olfactory receptors (ORs) as sensing elements instead of chemical sensors, biosensors would benefit the naturally optimized molecular recognition of odorants to develop a new generation of bioelectronic noses. The purpose of SPOT-NOSED European project was the development of nanobiosensors based on single ORs anchored between nanoelectrodes, to mimic the performances of natural olfactory system. Nanobiosensors arrays could then increase odorant sensitivity or widen the odorant detectionThe overall objective of the SPOT-NOSED (Single PrOTein NanObioSEnsor griD array) project was to explore the possibility to develop an olfactory nanobiosensor array based on the electrical properties of single olfactory receptors. The olfactory nanobiosensor array would integrate elementary nanobiotransducers, each of which consisting of a set of functionalised nanoelectrodes with an olfactory receptor monolayer anchored on them. The nanoelectrode would be in charge of detecting electrically any conformational or chemical change in the olfactory receptors when these receptors bind a given odorant molecule (ligand). The array structure would increase the sensitivity and specificity of the whole olfactory sensor. In order to reach the main objective of the project, advances needed to be performed in a variety of fields such as nanotechnologies, nanoelectronics, surface functionalization, expression, grafting, and electrical modelization of olfactory receptors. It required a very strong interaction within the Consortium, between research groups from very different fields, located in Spain, Italy, and France. Most results reported in the present paper were obtained within the framework of this project. Project website: spectrum. ORs were expressed in yeasts plasmic membrane, and their functionality tested in whole yeasts. Then, nanosomes bearing the ORs were prepared from S. cerevisiae, and Surface Plasmon Resonance was performed on nanosomes for quantitative evaluation of OR response to odorant stimulation. ORs retain full activity and discrimination power in immobilized nanosomes, thus allowing their use in the fabrication of the nanobiosensors. Nanoelectrodes were fabricated using conventional photolithography and focused ion beam milling, with sizes in adequation with the nanosomes. ORs borne by nanosomes were specifically immobilized onto conducting substrates via mixed Self Assembled Monolayers, neutravidin and specific antibody to the ORs. The process was optimized by microcontact printing, and the anchored nanovesicles visualized by Atomic Force Microscopy. A transimpedance preamplifier suited for low-noise wide-bandwidth measurements was designed to be directly connected to the nanoelectrodes. Electrochemical Impedancemetric Spectroscopy detected significant changes upon electrodes functionalization, grafting of ORs carried by nanosomes, and ORs c...

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Towards the Realization of Nanobiosensors Based on G‐protein‐coupled Receptors

Abstract: The sections in this article are Introduction Preparation and Immobilization of GPCRs on Functionalized Surfaces Signal Techniques Theoretical Approach The Impedance Network Model Equilibrium Fluctuations Conclusions … Show more

Cited by 12 publications

References 85 publications

A network model to correlate conformational change and the impedance spectrum of single proteins

A network model to correlate conformational change and the impedance spectrum of single proteins

Immobilization of Olfactory Receptors Carried by Nanosomes onto a Gold Sensor Surface

Nanobiosensors based on individual olfactory receptors

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