Dynamics and morphology of the in vitro polymeric form of elongation factor Tu from Escherichia coli.
Elongation factor Tu from Escherichia coli is known to polymerize at slightly acidic pH and low ionic strength. The structure and dynamics of these aggregates have been examined using imaging and spectroscopic methodologies. Electron microscopy provides evidence for two-dimensional sheets and bundled filaments of EF-Tu, whereas fluorescence microscopy of EF-Tu covalently labeled with tetramethylrhodamine isothiocyanate showed highly branched polymers of EF-Tu several microns in diameter. These polymers were studied using quasi-elastic light scattering to determine the evolution of the translational diffusion coefficient during the polymerization process. The rotational dynamics of the aggregate were investigated using phosphorescence anisotropy of EF-Tu covalently labeled with erythrosin isothiocyanate. A high infinite-time anisotropy was observed, suggesting a lack of motion or entanglement of EF-Tu polymers. A sub-microsecond motion which was slowed in the presence of glycerol may be due to local flexibility of the polymers. The possible relevance of polymeric EF-Tu to its function in vivo is discussed.
Department of Biochemistry and Biophysics, University of Hawaii, Honolulu 96822, USA.
M K
MK
Helms
G
G
Marriott
W H
WH
Sawyer
D M
DM
Jameson
D016880:Anisotropy; D004926:Escherichia coli / Q000737:chemistry*; D005990:Glycerol / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008027:Light; D008163:Luminescent Measurements; D008856:Microscopy, Fluorescence; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry*; D012542:Scattering, Radiation
D011108:Polymers; D010444:Peptide Elongation Factor Tu; D005990:Glycerol