Sensors: Biosensor Arrays from Intact Receptor Proteoliposomes Immobilized onto Surfaces

Couzis， Alexander等人在《传感器：来自固定在表面的完整受体蛋白脂体的生物传感器阵列》中指出，参与细胞分子传感和信号转导的主要受体家族是G蛋白偶联膜受体（gpcr），它们负责检测细胞外分子信号并通过偶联异三聚体G蛋白转导信号。GPCRs可以感知广泛的配体，包括神经递质、气味分子和生长因子。因此，GPCR传感器分子机制是用于生物传感器设计的自然系统。这项研究的目的是开发由gpcr组成的仿生传感器阵列。使用gpcr作为传感元件的主要限制是这些受体需要一个特殊的脂质双分子层膜环境来维持体外活性。在此方向上，先前的研究已经将GPCR结合到表面的平面双层结构中，以满足维持GPCR活性的脂质环境的要求。这项资助的研究方法是基于将完整的gpcr脂质体（蛋白脂质体）固定在具有控制终端功能域的纳米结构表面上。这些脂质体将在脂质双分子层中包含膜蛋白受体，更重要的是，它们还将G蛋白及其下游靶点如磷脂酶C包封在脂质体的核心。这种方法通过利用细胞的自然应用机制来显著提高GPCR阵列的灵敏度，因为它可能被封装在蛋白质脂质体内。原则上，该设计可以使用微流控概念进行放大，以探索配体、受体、G蛋白及其下游效应物的多种组合中的相互作用。该团队由来自纽约城市学院的研究人员组成，他们在生物物理学、生物工程、界面工程、生物化学、化学工程和遗传学方面有着良好的合作记录。这个团队的目的是发展科学和必要的技术来实现这样的设计，并在此过程中为研究生、本科生和研究生提供独特的、多学科的教育体验。此外，这提供了一个独特的机会，将研究生和本科生在表面科学（NSF 9972892）和软材料（NSF 0221589）方面的培训工作与美国国立卫生研究院（RCMI， MARC, &amp； RISE）资助的生物化学培训工作结合起来，否则这些培训工作将彼此独立地发挥作用。所有的合作伙伴都参与了上述一项或多项工作。这些培训工作将提供额外的资源，如本科生研究经费和一年级研究生经费。此外，该提案将利用最近从美国国家科学基金会和国防部获得基础设施资金的成功努力。

ABSTRACT Couzis, Alexander et alCUNY City College"Sensors: Biosensor Arrays from Intact Receptor Proteolipsomes Immobilized onto Surfaces"The dominant receptor family involved in molecular sensing and signal transduction by the cell are the G protein-coupled membrane receptors (GPCRs) which are responsible for detecting extracellular molecular signals and transducing the signals via the coupled heterotrimeric G protein. GPCRs sense an expansive range of ligands, including neurotransmitters, odorant molecules and growth factors. The GPCR sensor molecular machinery is therefore a natural system to be used in the design of biosensors. The aim of this research grant is to develop biomimetic sensor arrays composed of GPCRs. The primary limitation with using GPCRs as a sensing element is that these receptors require a specialized lipid bilayer membrane environment to maintain activity ex vivo. Previous efforts in this direction have incorporated the GPCRs into planar bilayer structures on surfaces in order to fulfill the requirement of a lipid environment to maintain GPCR activity. The research approach of this grant is based on the immobilization of intact GPCR-liposomes (proteoliposomes) onto nanostructured surfaces with domains of controlled terminal functionality. These liposomes will contain the membrane protein receptors in the lipid bilayer and, more importantly, also encapsulate G proteins and their downstream targets such as phospholipase C, in the core of the liposome. This approach stands to significantly enhance the senstivity of GPCR arrays by harnesing the natural applification mechanisms of the cell due to the potential for encapsulation inside the proteoliposome. In principle, the design can be scaled-up using microfluidic concepts to explore the interactions in a multitude of combinations of ligand, receptor, G proteins and their downstream effectors.The team consists of investigators with an established record of collaboration from the City College of New York, combining backgrounds in biophysics, bioengineering, interfacial engineering, biochemistry, chemical engineering, and genetics. It is the intention of this team to develop the science and required technology to achieve such a design and along the way provide a unique, multidisciplinary educational experience for the graduate, undergraduate, and post-graduate students that will be involved.Furthermore, this provides a unique opportunity to interface graduate and undergraduate training efforts in surface science (NSF 9972892) and soft materials (NSF 0221589) already funded by the NSF with biochemistry training efforts funded by the NIH (RCMI, MARC, & RISE) that would otherwise function independently of one another. All the co-PIs are involved in one or more of the above mentioned efforts. These training efforts will provide additional resources such as undergraduate research funding and first-year graduate student funding. In addition, this proposal will take advantage of the recent successful efforts to secure infrastructure funding from the NSF and the DoD.