Ian Wark Research Institute, ARC Special Research Centre, Mawson Lakes Campus, University of South Australia, Mawson Lakes, South Australia 5095, Australia.
Biomaterials. 2011 Apr;32(11):2786-96. doi: 10.1016/j.biomaterials.2010.12.030. Epub 2011 Feb 3.
In this investigation we report the synthesis of optically coupled hybrid architectures based on a new biomimetic fluorescent protein rec1-resilin and nanometer-scale gold nanoparticles (AuNPs) in a one-step method using a non-covalent mode of binding protocol. The presence of uniformly distributed fluorophore sequences, -Ser(Thr)-Tyr-Gly- along the molecular structure of rec1-resilin provides significant opportunity to synthesize fluorophore-modified AuNPs bioconjugates with unique photophysical properties. The detailed analyses of the AuNP-bioconjugates, synthesized under different experimental conditions using spectroscopic, microscopic and scattering techniques demonstrate the organizational pathways and the electronic and photophysical properties of the developed AuNP-rec1-resilin bioconjugates. The calculation of the bimolecular quenching constant using the Stern-Volmer equation confirms that the dominant mechanism involved in quenching of fluorescence of rec1-resilin in the presence of AuNP is static. Photoacoustic infrared spectroscopy was employed to understand the nature of the interfacial interaction between the AuNP and rec1-resilin and its evolution with pH. In such bioconjugates the quenched emission of fluorescence by AuNP on the fluorophore moiety of rec1-resilin in the immediate vicinity of the AuNP has significant potential for fluorescence-based detection schemes, sensors and also can be incorporated into nanoparticle-based devices.
在这项研究中,我们报告了一种新的仿生荧光蛋白 rec1-resilin 和纳米级金纳米粒子 (AuNPs) 的光学偶合杂化结构的合成,该方法采用非共价键结合方案一步法合成。rec1-resilin 分子结构中均匀分布的荧光序列 -Ser(Thr)-Tyr-Gly-为合成具有独特光物理性质的荧光标记 AuNPs 生物缀合物提供了重要机会。使用光谱、显微镜和散射技术对在不同实验条件下合成的 AuNP-生物缀合物进行的详细分析,证明了所开发的 AuNP-rec1-resilin 生物缀合物的组织途径和电子和光物理性质。使用 Stern-Volmer 方程计算双分子猝灭常数证实,在 AuNP 存在下,rec1-resilin 荧光猝灭的主要机制是静态的。光声红外光谱用于了解 AuNP 和 rec1-resilin 之间的界面相互作用的性质及其随 pH 值的演变。在这种生物缀合物中,AuNP 对 rec1-resilin 荧光团部分的荧光猝灭具有在 AuNP 附近的显著荧光基于检测方案、传感器的潜力,并且可以被整合到基于纳米粒子的设备中。