recombinant minimalist spider wrapping silk proteins capable of native-like fiber formation重组简约蜘蛛包装丝绸蛋白质类原生纤维形成的能力.pdfVIP

Recombinant Minimalist Spider Wrapping Silk Proteins Capable of Native-Like Fiber Formation 1,2 2,3 1 2 Lingling Xu , Jan K. Rainey , Qing Meng *, Xiang-Qin Liu * 1 Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai, P.R. China, 2 Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada, 3 Department of Chemistry, Dalhousie University, Halifax, Nova Scotia, Canada Abstract Spider silks are desirable biomaterials characterized by high tensile strength, elasticity, and biocompatibility. Spiders produce different types of silks for different uses, although dragline silks have been the predominant focus of previous studies. Spider wrapping silk, made of the aciniform protein (AcSp1), has high toughness because of its combination of high elasticity and tensile strength. AcSp1 in Argiope trifasciata contains a 200-aa sequence motif that is repeated at least 14 times. Here, we produced in E. coli recombinant proteins consisting of only one to four of the 200-aa AcSp1 repeats, designated W to W . We observed that purified W , W and W proteins could be induced to form silk-like fibers by shear 1 4 2 3 4 forces in a physiological buffer. The fibers formed by W4 were ,3.4 mm in diameter and up to 10 cm long. They showed an average tensile strength of 115 MPa, elasticity of 37%, and toughness of 34 J cm23. The smaller W2 protein formed fewer fibers and required a higher protein concentration to form fibers, whereas the smallest W1 protein did not form silk-like fibers, indicating that a minimum of two of the 200-aa repeats was required for fiber formation. M