NER: Two-Dimensional Sol Gel Protein Imprinting

NER：二维溶胶凝胶蛋白质印迹

• 批准号: 0404262
• 负责人: David Britt
• 金额: $ 10万
• 依托单位: Utah State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0404262&HistoricalAwards=false
• 关键词: NER; Two; Dimensional; Sol; Gel

Britt0404262This research aims to develop a two-dimensional imprint and molecular vise method for creating nanoscale sol-gel membranes for protein recognition. In-plane imprint sites are formed in floating monolayers of polymerizable alkyl-silanes co-spread with functionalized "target" silanes and poly(ethylene glycol) (PEG) bearing lipids. The lateral diffusivity of monolayer components facilitates a matching of amphiphile functional groups with residues on the adsorbing proteins through complementary interactions (hydrogen bonding, electrostatic), preserved upon monolayer polymerization. The Langmuir monolayer technique allows functional group density, lateral mobility, polymerization rate, gelled film morphology, PEG conformation, and protein insertion to be precisely controlled. Contraction of the imprinted monolayer upon alkyl-silane hydrolysis and condensation accompanied by barrier compression adds a "molecular vise" aspect to imprinting, where PEG chains that initially explore a large area in the "mushroom" phase are forced into the more ordered and conformationally restricted "brush" phase. This induced assembly of PEG bearing amphiphiles around proteins adsorbed to target silane head-groups builds binding pockets, while weakly adsorbed proteins (poor imprints) are squeezed out, improving binding site affinity and homogeneity. The PEG brush layer surrounding the in-plane imprints also reduces non-imprint mediated protein re-binding. The proposed collaborative research will help establish a nanoscience based partnership between the Biological Engineering department at Utah State University and the Bioengineering and Metallurgical Engineering departments at the University of Utah, leading to infrastructure enhancement and cooperative graduate and undergraduate student involvement from both universities

Britt 0404262本研究旨在开发一种二维印迹和分子虎钳方法，用于制备用于蛋白质识别的纳米溶胶-凝胶膜。平面内压印位点形成在与官能化“目标”硅烷和聚（乙二醇）（PEG）承载脂质共铺展的可聚合烷基硅烷的浮动单层中。单层组分的横向扩散性有利于通过互补相互作用（氢键、静电）使两亲性官能团与吸附蛋白上的残基匹配，在单层聚合后保留。朗缪尔单层技术允许官能团密度、横向流动性、聚合速率、凝胶膜形态、PEG构象和蛋白质插入被精确控制。在烷基硅烷水解和缩合伴随着屏障压缩的印迹单层的收缩增加了一个“分子虎钳”方面的印迹，其中PEG链，最初探索一个大面积的“蘑菇”相被迫进入更有序和构象限制的“刷”相。这种在吸附到目标硅烷头基的蛋白质周围的PEG承载两亲物的诱导组装建立了结合口袋，而弱吸附的蛋白质（不良印记）被挤出，改善了结合位点亲和力和均匀性。包围平面内印记的PEG刷层也减少了非印记介导的蛋白质再结合。拟议的合作研究将有助于在犹他州州立大学生物工程系与犹他州大学生物工程和冶金工程系之间建立基于纳米科学的伙伴关系，从而增强基础设施，并促进两所大学研究生和本科生的合作参与