An analysis of FtsZ assembly using small angle X-ray scattering and electron microscopy.
Small angle X-ray scattering (SAXS) was used for the first time to study the self-assembly of the bacterial cell division protein, FtsZ, with three different additives: calcium chloride, monosodium glutamate and DEAE-dextran hydrochloride in solution. The SAXS data were analyzed assuming a model form factor and also by a model-independent analysis using the pair distance distribution function. Transmission electron microscopy (TEM) was used for direct observation of the FtsZ filaments. By sectioning and negative staining with glow discharged grids, very high bundling as well as low bundling polymers were observed under different assembly conditions. FtsZ polymers formed different structures in the presence of different additives and these additives were found to increase the bundling of FtsZ protofilaments by different mechanisms. The combined use of SAXS and TEM provided us a significant insight of the assembly of FtsZ and microstructures of the assembled FtsZ polymers.
Kuchibhatla
Anuradha
A
School of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai, India 400 076.
Abdul Rasheed
A S
AS
Narayanan
Janaky
J
Bellare
Jayesh
J
Panda
Dulal
D
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002122:Calcium Chloride / Q000737:chemistry; D002455:Cell Division; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000378:metabolism; D003637:DEAE-Dextran / Q000737:chemistry; D018698:Glutamic Acid / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron / Q000379:methods; D015233:Models, Statistical; D010316:Particle Size; D011108:Polymers / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D012970:Sodium Glutamate / Q000737:chemistry; D014965:X-Rays
D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D011108:Polymers; D011994:Recombinant Proteins; D018698:Glutamic Acid; D003637:DEAE-Dextran; D002122:Calcium Chloride; D012970:Sodium Glutamate