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Molecular docking demonstrating that the X-ray crystal structure of the ZipA GD, fits well into the smaller lobe in all dimensions of the 3D structures in all three models, determined using orthogonal structural techniques. This suggests that this small lobe is the ZipA GD with the rest of the protein being internal to the SMALP. Analysis of the final structure shows that the ZipA GD is separated from the lipid and SMA by a thinner region of density, likely to be the extended domain PQ, which forms a flexible tether between the ZipA GD and the 1 TMH fixed into the membrane.
The SMALP-ZipA:FtsZ complex shows a particle that is significantly larger than SMALP-ZipA alone. This structure is also bi-lobular with one lobe of similar size to that attributed to the lipid and SMA in the SMALP-ZipA structure and a second lobe that is larger. The larger lobe is consistent with models fitting a ZipA GD:FtsZ complex into this density. A static analysis of the structure shows that the area of the particle that correlates with unstructured region of ZipA remains relatively unchanged indicating that there is no significant conformational change in the presence of bound FtsZ.
S.C.L., T.R.D., D.I.R. and A.R. wrote the manuscript. S.C.L., A.T., C.T. and K.J.E. performed SAXS and SANS experiments analysed the data. S.C.L. and C.B. D.I.R and A.R FtsZ production, negative stain analysis with polymerized SMALP-ZipA, SMALP-ZipA:FtsZ FtsZ polymerisation, circular dichroism data analysis. S.C.L., Y.L., R.P. and M.J. Expression, purification of ZipA, AUC, co-precipitation of ZipA with FtsZ. S.C.L., R.C., C.J.I.S. and R.F. Cryo-EM and 3D reconstruction of SMALP-ZipA and SMALP-ZipA:ftsZ. S.C.L., B.H. and S.H. carried out the FFEA concepts and initial ideas and modelling. B.H. produced carried out the simulations, prepared models and movies. S.C.L. prepared all of the figures.
Jaeger, Kristin L., Konrad C. Hafen, Jason B. Dunham, Ken M. Fritz, Stephanie K. Kampf, Theodore B. Barnhart, Kendra E. Kaiser, Roy Sando, Sherri L. Johnson, Ryan R. McShane, and Sarah B. Dunn. 2021. "Beyond Streamflow: Call for a National Data Repository of Streamflow Presence for Streams and Rivers in the United States" Water 13, no. 12: 1627. 350c69d7ab