K(+)-ribosome interactions determine the large enhancements of 39K NMR transverse relaxation rates in the cytoplasm of Escherichia coli K-12.
As a probe of physical chemical properties of the intracellular environment, we measured 39K NMR transverse relaxation rates in concentrated cell slurries of Escherichia coli K-12 grown in minimal medium over a range of osmolarities (from 0.1 to 1.0 OsM) and after plasmolysis. The 39K transverse relaxation at a resonance frequency of approximately 18.67 MHz is biexponential under all conditions, and 100% of the expected signal intensity is detected. Both components of the 39K NMR transverse relaxation are very fast, and the difference between the fast and slow relaxation rates is very large compared to previous measurements on 23Na and 39K in protein and nucleic acid solutions in vitro. The 39K transverse relaxation rates decrease as the osmolarity of the growth media increases but increase dramatically when cells grown in 0.1 OsM media are plasmolyzed at 1.0 OsM. The homogeneous nature and the 100% visibility of the 39K signal indicate the existence of fast exchange among the multiple, magnetically distinguishable populations of 39K which probably exist in the cytoplasm. The absence of static quadrupolar splitting of the cytoplasmic 39K signal (as indicated by a single peak in the spectrum) indicates that the cytoplasm, as probed by 39K NMR, behaves like a concentrated but isotropic nucleic acid solution rather than an anisotropic nucleic acid liquid crystal. To understand the origins of the striking NMR relaxation behavior of 39K in viable cells, we have investigated NMR transverse relaxation rates of 39K (and also 23Na and 35Cl) in E. coli 50S and 70S ribosome solutions in vitro. At concentrations of ions and of ribosomes that to the extent possible mimic those of the cytoplasm of E. coli, we find that 39K, 23Na, and 35Cl transverse relaxation rates all exhibit biexponential behavior, and 39K and 23Na exhibit the large magnitudes and the large difference between the slow and the fast relaxation rates observed in viable cells. These polyanionic ribosome solutions are the only in vitro model system discovered to date that exhibits 39K transverse relaxation rates comparable to those in viable cells. We conclude that K(+)-ribosome interactions are the dominant source of the NMR properties of K+ in E. coli.(ABSTRACT TRUNCATED AT 400 WORDS)
Guttman
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HJ
Department of Chemistry, University of Wisconsin-Madison 53706.
Cayley
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Li
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Anderson
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Record
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D003593:Cytoplasm; D004926:Escherichia coli / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009038:Motion; D011188:Potassium / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D011188:Potassium