Assembly Mechanism of Recombinant Proteins

研究人员已经能够对具有新特性的蛋白质材料进行基因工程改造。例如，丝弹性蛋白样蛋白（SELP）已经通过将多肽序列融合成单一蛋白质而合成，所述多肽序列源自具有上级机械强度的天然丝，并且所述人弹性蛋白具有显著的弹性。然而，重组蛋白，如SELP，仍然不能组装成强大的微/纳米生物医学结构通过？绿色？流程.与此相反，蜘蛛可以在室温、低压和以水为溶剂的条件下纺出上级强度的丝纤维。这个建议的目的是了解下重组蛋白的组装机制？绿色？条件下组装的蛋白质纤维的结构和力学性能进行表征。特别地，微流体装置将用于研究蛋白质溶液变量（例如，pH、盐浓度）、制造变量（例如，流速）和蛋白质序列对SELP的受控组装和纤维形成的影响。组装的SELP纤维的机械性能将被表征和数学建模。此外，SELP光纤的二级结构将使用傅里叶变换红外拉曼光谱进行分析。拟议的研究将提供必要的第一步发展？绿色？高性能蛋白质纤维的制造工艺，用于生产各种生物医学应用。从研究结果将被整合到合并的本科生和研究生课程题为微观生物力学。 进行研究的本科生和研究生，包括代表性不足的少数民族和妇女，将接受生物力学，纳米技术和生物材料等新兴领域的培训。

Researchers have been able to genetically engineer protein-based materials with novel properties. For instance, silk-elastin-like proteins (SELPs) have been synthesized by fusing polypeptide sequences, derived from native silk of superior mechanical strength, and human elastin, which is remarkably resilient, into single proteins. However, recombinant proteins, like SELPs, still cannot be assembled into robust micro/nanoscale biomedical structures via ?green? processes. In contrast, spiders can spin silk fibers of superior strength at ambient temperatures, low pressures, and with water as the solvent. The objective of this proposal is to understand the assembly mechanism of recombinant proteins under ?green? conditions and to characterize the structure and mechanical properties of assembled protein fibers. In particular, a micro-fluidic device will be used to investigate the influence of protein solution variables (e.g., pH, salt concentrations), fabrication variables (e.g., flow rates), and protein sequences on the controlled assembly and fiber formation of SELPs. The mechanical properties of assembled SELP fibers will be characterized and mathematically modeled. In addition, the secondary structures of SELP fibers will be analyzed using Fourier transform infrared Raman spectroscopy. The proposed research will provide the necessary first step toward the development of ?green? fabrication processes for producing high-performance protein fibers for a variety of biomedical applications. Results from the research will be integrated into the combined undergraduate and graduate course entitled Micro-Biomechanics. Undergraduate and graduate students performing the research, including underrepresented minorities and women, will receive training in the emerging areas of biomechanics, nanotechnology, and biomaterials.