PMID: 698203Sep 5, 1978Paper

Fluorotyrosine M13 coat protein: fluorine-19 nuclear magnetic resonance study of the motional properties of an integral membrane protein in phospholipid vesicles

Biochemistry
D S HagenB D Sykes

Abstract

We have prepared in vivo a fluorotyrosyl derivative of M13 coat protein and have incorporated it at high levels in small phospholipid vesicles, using a urea-cholate dialysis procedure. 19F nuclear magnetic resonance experiments at 254 MHz with this system indicate a T1 of 0.32 s and line width of 300 Hz. The observed line width increases dramatically below the gel to liquid-crystalline transition temperature for the lipid, indicating that the probe is sensitive to the phase state of the bilayer. Neclear Overhauser enhancement and field dependence of line width were used to establish the relative contributions of dipolar interactions and chemical-shift anisotropy to the observed T1 and line width. From this relaxation data, we have constructeda model for the motional properties of the protein in the lipid bilayer. This model is characterized by correlation times for rotation about the alphabeta and betagamma bonds of the two tyrosyl residues of 2 x 10(-8) and k x 10(-9) s, respectively. Rapid intermolecular dipole-dipole interactions are required to account for theestimated dipolar contribution to T1. A reasonable model for these interactions is that lipid methylene protons are involved in relaxation of the fluorine probes (whic...Continue Reading

References

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Citations

Apr 5, 1984·Biochimica Et Biophysica Acta·M J Wilson, D L Hatfield
Apr 27, 1987·Biochimica Et Biophysica Acta·M K Jain, D Zakim
Apr 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·M C LoewenH G Khorana
May 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·M A JaremaJ H Miller
Jan 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·C M GallS J Opella
Sep 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·H KoideT Miyazawa
Mar 4, 2014·Progress in Biophysics and Molecular Biology·D A MarvinS K Straus
Jan 29, 1980·Biochemical and Biophysical Research Communications·T A Cross, S J Opella
Aug 13, 2013·Current Opinion in Structural Biology·Tatiana DidenkoKurt Wüthrich
May 29, 1979·Biochemistry·L M SmithH M McConnell
Feb 10, 1987·Biochemistry·G C LeoS J Opella
May 9, 1990·Biochimica Et Biophysica Acta·H H de JonghD Marsh
Dec 1, 1986·Microbiological Reviews·I Rasched, E Oberer

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