SDS-PAGE profile of the FtsZ polymer pelleting assay for MsFtsZ polymerisation and its quantitation.

<p>The SDS-PAGE profile of the sample in the pellet (<b>A</b>) and in the supernatant (<b>B</b>). Its quantitation is given as the pellet/supernatant ratio in (<b>C</b>). (<b>A & B</b>) Lanes: 1 & 9, MsFtsZ + GTP; 2 & 10, MsFtsZ without GTP; 3 & 11, MsFtsZ + MsNDK-Wt + GDP + ATP; 4 & 12, MsFtsZ + MsNDK-H117Q + GDP + ATP; 5 & 13, MsFtsZ + MtNDK-Wt + GDP + ATP; 6 & 14, MsFtsZ + MtNDK-H117Q + GDP + ATP; 7 & 15, GDP-precharged MsFtsZ + MsNDK-Wt + ATP; 8 & 16, GDP-precharged MsFtsZ + MtNDK-Wt + ATP. (<b>C</b>) Quantitations of the protein as the pellet/supernatant ratio in the reactions are represented by the bars, which are described in the figure itself. (<b>D</b>) The SDS-PAGE profile of the samples in the supernatant and pellet from MsFtsZ polymerisation in the absence of exogenous GDP. (<b>E</b>) Its quantitation given as the pellet/supernatant ratio. Lanes in (<b>D</b>): 1, MsFtsZ + GTP (Sup); 2, MsFtsZ—GTP (Sup); 3, MsFtsZ + MsNDK-Wt + ATP (Sup); 4, MsFtsZ + MsNDK-H117Q + ATP (Sup); 5, Marker; 6, MsFtsZ + GTP (Pellet); 7, MsFtsZ—GTP (Pellet); 8, MsFtsZ + MsNDK-Wt + ATP (Pellet); 9, MsFtsZ + MsNDK-H117Q + ATP (Pellet). (<b>E</b>) Quantitations of the protein as the pellet/supernatant ratio in the reactions are represented by the bars, which are described in the figure itself. (<b>F-H</b>) Comparison of TEM images (105k magnification) of FtsZ polymers formed from polymerisation of MsFtsZ triggered by MsNDKwt and MsNDKmut in the absence and presence of exogenous GDP. (<b>F</b>) MsFtsZ polymers from MsZ + NDKwt + ATP reaction; (<b>G</b>) MsFtsZ polymers from MsZ + NDKwt + GDP + ATP reaction; (<b>H</b>) MsFtsZ polymers from MsZ + NDKmut + ATP reaction. Images were captured in FEI Tecnai BioTWIN EM at 105k magnification.