RESUMO
From roughly 1985 through the start of the new millennium, the cutting edge of solution protein nuclear magnetic resonance (NMR) spectroscopy was to a significant extent driven by the aspiration to determine structures. Here we survey recent advances in protein NMR that herald a renaissance in which a number of its most important applications reflect the broad problem-solving capability displayed by this method during its classical era during the 1970s and early 1980s.
Assuntos
Ressonância Magnética Nuclear Biomolecular/métodos , Proteínas/química , Animais , Descoberta de Drogas , História do Século XX , História do Século XXI , Humanos , Modelos Moleculares , Ressonância Magnética Nuclear Biomolecular/história , Conformação Proteica , Mapeamento de Interação de Proteínas/história , Mapeamento de Interação de Proteínas/métodos , Proteínas/metabolismoRESUMO
In 1996, the Wickens and the Kuhl labs developed the yeast three-hybrid system independently. By expressing two chimeric proteins and one chimeric RNA molecule in Saccharomyces cerevisiae, this method allows in vivo monitoring of RNA-protein interactions by measuring the expression levels of HIS3 and LacZ reporter genes. Specific RNA targets have been used to characterize unknown RNA binding proteins. Previously described RNA binding proteins have also been used as bait to select new RNA targets. Finally, this method has been widely used to investigate or confirm previously suspected RNA-protein interactions. However, this method falls short in some aspects, such as RNA display and selection of false positive molecules. This review will summarize the results obtained with this method from the past 15years, as well as on recent efforts to improve its specificity.
Assuntos
Proteínas de Ligação a RNA/metabolismo , RNA/metabolismo , Técnicas do Sistema de Duplo-Híbrido/normas , Animais , Biblioteca Gênica , História do Século XX , História do Século XXI , Humanos , Ligação Proteica , Mapeamento de Interação de Proteínas/história , Mapeamento de Interação de Proteínas/métodos , Mapeamento de Interação de Proteínas/normas , Saccharomyces cerevisiae , Sensibilidade e Especificidade , Técnicas do Sistema de Duplo-Híbrido/históriaRESUMO
Today, it is widely appreciated that protein-protein interactions play a fundamental role in biological processes. This was not always the case. The study of protein interactions started slowly and evolved considerably, together with conceptual and technological progress in different areas of research through the late 19th and the 20th centuries. In this review, we present some of the key experiments that have introduced major conceptual advances in biochemistry and molecular biology, and review technological breakthroughs that have paved the way for today's systems-wide approaches to protein-protein interaction analysis.
Assuntos
Bioquímica/história , Mapeamento de Interação de Proteínas/história , Proteínas/química , Proteínas/metabolismo , Biologia de Sistemas/história , Animais , Bioquímica/métodos , História do Século XIX , História do Século XX , História do Século XXI , Humanos , Mapeamento de Interação de Proteínas/métodos , Transdução de Sinais , Biologia de Sistemas/métodosRESUMO
Some historical background is given for appreciating the impact of the empirical construct known as the cellular protein-protein interactome, which is a seemingly de novo entity that has arisen of late within the context of postgenomic systems biology. The approach here builds on a generalized principle of "fuzziness" in protein behavior, proposed by Tompa and Fuxreiter.(1) Recent controversies in the analysis and interpretation of the interactome studies are rationalized historically under the auspices of this concept. There is an extensive literature on protein-protein interactions, dating to the mid-1900s, which may help clarify the "fuzziness" in the interactome picture and, also, provide a basis for understanding the physiological importance of protein-protein interactions in vivo.
