![]() ![]() (SI 2.10) Dynamic light scattering (DLS) – 50 μL of an assembly sample was used for size determination using a Malvern Zetasizer and a quartz cuvette (ZEN2112, Malvern). Size measurements using DLS were performed to determine assembly formation/disassembly. Disassembly was performed by adding 4.8μg of YopH phosphatase into the 150μL reaction mixture after assembly formation occurred. After phosphorylating, AtzCM1 was added to a final 2μM concentration. In a final reaction volume of 150μL, 3μM AtzAM1 was mixed into 1X Kinase Activity Buffer (4mM MgCl2, 2.5mM MnCl2, 0.25mM DTT, 5mM MOPS, 2.5mM glycerol-2-phosphate, 1mM EGTA, 400nM EDTA, pH 7.6), 2.5 mM MnCl2,HNG, 2 mM ATP, 800ng Src kinase, and incubated for 7 – 16 hr at 25☌ for phosphorylation to occur. (SI 2.9) Phosphorylation, assembly formation, and disassembly – The phosphorylation protocol was based upon Src kinase activity assay by Sigma (Catalog # S1076). Distributions shown are representative of other traces in the sample. Histogram illustrates distribution of sizes found on heatmap. Value shown is average of two physical samples. (A) Heat map showing volume-weighted mean size of particles found from 50-3000 nM pY-AtzA and 50-2000 nM AtzC-SH2. ![]() Average size of particle formed by pY-AtzA and super-binder AtzC-SH2. Article: Stimulus-responsive Self-Assembly of Protein-Based Fractals by Computational Designįigure: Fig. ![]()
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