Pompon D, Laisné A
Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, UPR2167, Avenue de la Terrasse, 91190 Gif-sur-Yvette, France.
Biochem Soc Trans. 2007 Jun;35(Pt 3):495-7. doi: 10.1042/BST0350495.
Different semi-synthetic PDNAs (protein-DNA complexes), which encompass a protein core engineered from the cytochrome b(5) scaffold, an embedded tuneable redox cofactor, a synthetic linker and a large oligonucleotide, were designed, synthesized and purified to homogeneity. These building blocks can be reversibly attached to Ni-DOGS [1,2-dioleoyl-sn-glycero-3-[N(5-amino-1-carboxypentyl)iminodiacetic acid]succinyl]-doped supported membranes through a metal chelate bridge with the protein part and be polymerized in a fully controllable manner using a solid-phase synthesis strategy and a stepwise addition of suitable complementary oligonucleotides. The resulting structures could recreate a large range of regular distribution of patterned redox and absorbing centres separated by fully tuneable distances and geometry. Kinetic parameters for the self-assembly of building blocks were determined using SPRI (surface plasmon resonance imagery). Structures of resulting nano-objects were characterized using gel electrophoresis and single molecule approaches following decoration of assemblies with quantum dots.
设计、合成并纯化了不同的半合成蛋白质 - DNA 复合物（PDNAs），其包含由细胞色素 b(5) 支架工程改造而来的蛋白质核心、嵌入的可调谐氧化还原辅因子、合成接头和大的寡核苷酸，直至达到均一性。这些构建模块可通过与蛋白质部分形成的金属螯合桥，可逆地连接到掺杂了 Ni - DOGS（1,2 - 二油酰基 - sn - 甘油 - 3 - [N(5 - 氨基 - 1 - 羧基戊基)亚氨基二乙酸]琥珀酰）的支撑膜上，并使用固相合成策略和逐步添加合适的互补寡核苷酸以完全可控的方式进行聚合。所得结构能够重现大量具有规则分布的图案化氧化还原和吸收中心，这些中心之间的距离和几何形状可完全调谐。使用表面等离子体共振成像（SPRI）确定构建模块自组装的动力学参数。在用量子点修饰组装体后，使用凝胶电泳和单分子方法对所得纳米物体的结构进行表征。
