Designed DNA-DNA interactions are investigated for their ability to modulate protein packing within single crystals of mutant green fluorescent proteins (mGFPs) functionalized with a single DNA strand (mGFP-DNA). We probe the effects of DNA sequence, length, and protein-attachment position on the formation and protein packing of mGFP-DNA crystals. Notably, when complementary mGFP-DNA conjugates are introduced to one another, crystals form with nearly identical packing parameters, regardless of sequence if the number of bases is equivalent. DNA complementarity is essential, because experiments with non-complementary sequences produce crystals with different protein arrangements. Importantly, the DNA length and its position of attachment on the protein markedly influence the formation of and protein packing within single crystals. This work shows how designed DNA interactions can be used to influence the growth and packing in X-ray diffraction quality protein single crystals and is thus an important step forward in protein crystal engineering.

对设计的DNA-DNA相互作用进行了研究，以研究其在具有单条DNA链（mGFP-DNA）功能的突变绿色荧光蛋白（mGFPs）的单晶中调节蛋白质堆积的能力。我们探讨了DNA序列，长度和蛋白质附着位置对mGFP-DNA晶体形成和蛋白质堆积的影响。值得注意的是，当互补的mGFP-DNA共轭物相互引入时，无论碱基数是否相等，无论序列如何，晶体都具有几乎相同的堆积参数。DNA互补性至关重要，因为使用非互补序列进行的实验会产生具有不同蛋白质排列的晶体。重要的是，DNA长度及其在蛋白质上的附着位置显着影响单晶内蛋白质的形成和蛋白质堆积。