Theoretical Studies of Fluorescent Proteins

描述(申请人提供)：对绿色荧光(GF)和其他光活性蛋白质进行研究，重点是阐明光化学循环和质子转移的机制，以支持在这些重要的生化标记中观察到的复杂光谱和相关行为。对这一领域迅速增长的文献进行了研究，并选择了单分子和主体相的快速荧光和相关测量，在使用理论和计算方法的研究计划中进行了解释。注意GFP的突变变体、光活性黄色蛋白和最近开发的在特定蛋白质位置进行内部溶剂化的“非天然”氨基酸荧光探针的不同行为。GFP作为一种可克隆的非侵入性标记物，可以在活细胞中用作基因表达和蛋白质定位的报告，检测蛋白质相互作用，以及作为各种生理条件的细胞内传感器，这一公认的重要性表明，它的研究可以为更广泛地了解蛋白质的性质提供更广泛的基础。更好地理解GFP的详细工作原理可能有助于开发用于特定目的的新的改进的荧光标记物，包括检测从单细胞扫描水平到全身扫描水平的转移，以及其他潜在的诊断优势。

DESCRIPTION (provided by applicant): Studies are performed of green fluorescent (GF) and other photoactive proteins, with emphasis on elucidating the photochemical cycles and mechanisms of proton transfer thought to underlie the complex spectroscopic and related behaviors observed in these important biochemical markers. The rapidly growing body of literature in this area is studied, and selected fast-fluorescence and related measurements in both single-molecule and bulk phases interpreted in a research program employing theoretical and computational methods. Attention is addressed to differing behaviors of mutant variants of GFP, of photoactive yellow proteins, and of recently developed "non-natural" amino acid fluorescent probes of internal solvation at specific protein sites. The recognized importance of GFP as a cloneable 'non-invasive marker, which can be employed in living cells as a reporter of gene expression and protein localization, in the detection of protein-protein interactions, and as an intra-cellular sensor of various physiological conditions, suggests its study can provide a broadly-based introduction to the properties of proteins more generally. Better understanding of the detailed workings of GFP may contribute to development of new improved fluorescent markers for specified purposes, including detection of metastases from single-cell to whole-body scanning levels, among other potential diagnostic benefits.