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. 2020 Dec 7;10:21369. doi: 10.1038/s41598-020-78431-x

Figure 1.

Figure 1

(A–F) Negative stain EM of PaFtsZ assembly in different buffer conditions. (A,B) 3 and 5 µM PaFtsZ in HMK (50 mM HEPES, 100 mM KAc, 5 mM MgAc, pH 7.5). (C,D) 5 µM PaFtsZ in MMK (50 mM MES, 100 mM KAc, 5 mM MgAc, pH 6.5) at 1 min (C) and 5 min after adding GTP (D). (E) 5 µM PaFtsZ in MEK (50 mM MES, 100 mM KAc, 1 mM EDTA, pH 6.5). The arrow indicates a close circle. (F) 10 µM PaFtsZ in HEK (50 mM HEPES, 100 mM KAc, 1 mM EDTA, pH 7.5). The bar equals 200 nm and all EM images have the same magnification. (G,H) Assembly kinetics of 5 µM PaFtsZ measured by the light-scattering assay. PaFtsZ assembled mostly single filaments within 10 s in HMK buffer (G), but in MMK buffer, it is a two-stage assembly: single filaments assembled within 10 s, followed by a second increase in light scattering attributed to bundling (H). Assembly in MMK followed for a longer time and on a larger light scattering scale (H). It is worth mentioning that the intensity values between (G) and (H) are not comparable. We reduce the sensitivity of the detector to measure the bundling formation (H). (I) GTPase activity of PaFtsZ at different concentrations in HMK and MMK buffer. GTP is 0.5 mM.