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Tunable alignment of macromolecules by filamentous phage yields dipolar coupling interactions

Nature Structural Biology volume 5, pages 10651074 (1998) | Download Citation

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Abstract

Dipolar coupling interactions represent an extremely valuable source of long-range distance and angle information that was previously not available for solution structure determinations of macromolecules. This is because observation of these dipolar coupling data requires creating an anisotropic environment for the macromolecule. Here we introduce a new method for generating tunable degrees of alignment of macromolecules by addition of magnetically aligned Pf1 filamentous bacteriophage as a cosolute. This phage-induced alignment technique has been used to study 1H-1H, 1H-13C, and 1H-15N dipolar coupling interactions in a DNA duplex, an RNA hairpin and several proteins including thioredoxin and apo-calmodulin. The phage allow alignment of macromolecules over a wide range of temperature and solution conditions and thus represent a stable versatile method for generating partially aligned macromolecules in solution.

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Acknowledgements

This work was supported by grants from NIH (to A.P.) and a postdoctoral fellowship from the Leukemia Society of America (to M.R.H.). We thank A. Bax for providing the alignment tensor program, M.S. Friedrichs for modifications of the program, J. Wank for technical assistance, K.J. Addess and A.J. Wand for preparation of samples and M. Rance and D.S. Wuttke for valuable discussions.

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Affiliations

  1. Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, Colorado 80309-0215, USA.

    • Mark R. Hansen
    •  & Arthur Pardi
  2. Bristol-Myers Squibb Pharmaceutical Research Institute, P.O. Box 4000, Princeton, New Jersey 08543-4000, USA.

    • Luciano Mueller

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Correspondence to Arthur Pardi.

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DOI

https://doi.org/10.1038/4176

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