Combined analytical ultracentrifugation, light scattering and fluorescence spectroscopy studies on the functional associations of the bacterial division FtsZ protein

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• 作者：Begoñ ; a Monterroso^a ; ^{monterroso@cib.csic.es} ; Carlos Alfonso^a ; ^{carlosa@cib.csic.es} ; Silvia Zorrilla^b ; ^{silvia@iqfr.csic.es} ; Germá ; n Rivas^a ; ^{grivas@cib.csic.es}
• 关键词：Cc ; critical concentration of assembly ; CG-SLS ; composition gradient static light scattering ; DLS ; dynamic light scattering ; FCS ; fluorescence correlation spectroscopy ; GDP-FtsZ ; GDP bound FtsZ ; GMPCPP ; guanosine-5&prime ; - [(¦Á ; ¦Â)-methylene] triphosphate
• 刊名：Methods
• 出版年：2013
• 出版时间：1 March, 2013
• 年：2013
• 卷：59
• 期：3
• 页码：349-362
• 全文大小：1069 K

文摘

The combined application of different biophysical techniques - analytical ultracentrifugation, light scattering and fluorescence-based assays - to study the ligand-linked self-association and assembly properties of the cell division protein FtsZ from Escherichia coli is described. These reactions are thought to be important for the formation of the dynamic division ring that drives bacterial cytokinesis. In addition, the use of this orthogonal experimental approach to measure the interactions between FtsZ oligomers (GDP forms) and polymers (GTP forms) with two variants (a soluble form and a full-length protein incorporated in phospholipid bilayer nanodiscs) of the ZipA protein, which provides membrane tethering to FtsZ, is described as well. The power of a global analysis of the results obtained from complementary biophysical methods to discriminate among alternative self- and hetero-associating schemes and to propose a more robust description of the association reactions involved is emphasized. This orthogonal approach will contribute to complete our quantitative understanding of the initial events of bacterial division.