RESUMEN
Herein, we report progress toward the development of bioactive surfaces based on gamma-aminobutyric acid (GABA), a major neurotransmitter in the nervous system. Whereas immobilization techniques have focused largely on antibodies, enzymes, and receptors, to our knowledge, this is the first report of a prototype neurotransmitter-immobilized surface. Biosurfaces were assembled onto either mica or glass using passive adsorption of avidin and subsequent attachment of a derivatized form of GABA via a biotin-avidin affinity bond. Surface characterization of these prepared bimolecular surfaces was determined using atomic force microscopy in tapping mode. The data reveal that passive adsorption of avidin is uniformly dispersed and cluster densities can be controlled through the concentration of the avidin incubation solution. GABA tethered via biotin to these avidin surfaces displayed a unique surface topology; in addition, histograms of surface heights suggest two different types of molecular cluster populations. Functional assays were performed to test the biological activity of the synthesized GABA. Anti-GABA antibody directed to these bimolecular surfaces result in morphological topologies and histograms that indicate antibody-antigen binding. However, nonspecific anti-immunoglobulin G antibodies directed to these surfaces show low binding affinity. Taken together, the data support the idea that the synthesized surfaces are biofunctional.
Asunto(s)
Receptores de GABA/metabolismo , Ácido gamma-Aminobutírico/química , Adsorción , Silicatos de Aluminio , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Reacciones Antígeno-Anticuerpo , Avidina/química , Biotinilación , Vidrio , Ratones , Microscopía de Fuerza Atómica , Propiedades de Superficie , Ácido gamma-Aminobutírico/análogos & derivados , Ácido gamma-Aminobutírico/inmunología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
The cyclic amino acid surrogate 1 was designed to mimic the extended conformation of a peptide unit and to provide hydrogen bond donor and acceptor functions conducive to beta-sheet formation. A convenient synthesis of this unit and solution and solid-phase methods for its incorporation into an oligomer alternating with peptide units have been devised. The resulting "@-tides", as these oligomers have been designated, show a high propensity for self-association in comparison to oligopeptides; insights into the structure and dynamical properties of their antiparallel dimers have been obtained by NMR.