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BMC Biotechnol ; 5: 18, 2005 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-15960850

RESUMO

BACKGROUND: Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processes has been considerably more difficult. RESULTS: In this article, we describe a novel technology for generating cell membrane microarrays for performing high throughput biology. Our robotically-arrayed supported membranes are physiologically fluid, a critical property which differentiates this technology from other previous membrane systems and makes it useful for studying cellular processes on an industrialized scale. Membrane array elements consist of a solid substrate, above which resides a fluid supported lipid bilayer containing biologically-active molecules of interest. Incorporation of transmembrane proteins into the arrayed membranes enables the study of ligand/receptor binding, as well as interactions with live intact cells. The fluidity of these molecules in the planar lipid bilayer facilitates dimerization and other higher order interactions necessary for biological signaling events. In order to demonstrate the utility of our fluid membrane array technology to ligand/receptor studies, we investigated the multivalent binding of the cholera toxin B-subunit (CTB) to the membrane ganglioside GM1. We have also displayed a number of bona fide drug targets, including bacterial endotoxin (also referred to as lipopolysaccharide (LPS)) and membrane proteins important in T cell activation. CONCLUSION: We have demonstrated the applicability of our fluid cell membrane array technology to both academic research applications and industrial drug discovery. Our technology facilitates the study of ligand/receptor interactions and cell-cell signaling, providing rich qualitative and quantitative information.


Assuntos
Biotecnologia/métodos , Membrana Celular/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Análise Serial de Proteínas/métodos , Animais , Sítios de Ligação , Diferenciação Celular , Toxina da Cólera/química , Dimerização , Desenho de Fármacos , Indústria Farmacêutica , Gangliosídeo G(M1)/química , Antígenos de Histocompatibilidade Classe II/química , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/biossíntese , Ligantes , Bicamadas Lipídicas/química , Lipopolissacarídeos/metabolismo , Membranas/metabolismo , Camundongos , Camundongos Transgênicos , Ligação Proteica , Mapeamento de Interação de Proteínas , Robótica , Salmonella enterica/metabolismo , Transdução de Sinais , Linfócitos T/metabolismo , Tecnologia Farmacêutica
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