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NMR of Membrane Proteins in Membranes Francesca M. Marassi Burnham Institute for Medical Research

NMR of Membrane Proteins in Membranes Francesca M. Marassi Burnham Institute for Medical Research NIGMS Roadmap for Membrane Protein Structures March 27 2009, UC San Francisco. E. coli OmpX. Y. Pestis Ail. M. tuberculosis OmpATb. Complementary approaches to membrane proteins:

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NMR of Membrane Proteins in Membranes Francesca M. Marassi Burnham Institute for Medical Research

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  1. NMR of Membrane Proteins in Membranes Francesca M. Marassi Burnham Institute for Medical Research NIGMS Roadmap for Membrane Protein Structures March 27 2009, UC San Francisco

  2. E. coli OmpX. • Y. Pestis Ail. • M. tuberculosis OmpATb. • Complementary approaches to membrane proteins: • β-barrel bacterial outer membrane proteins. solution NMR in micelles solid-state NMR in lipid bilayers X-ray diffraction In crystals

  3. Marassi and Opella (2000) J. Magn. Reson. 144:150Wang et al. (2000) J. Magn. Reson. 144:162 Marassi (2001) Biophys. J. 80:994

  4. Ail/Lom bacterial outer membrane proteins. • adhesion to host epithelial cells • resistance to complement-mediated killing • pathogenesis

  5. Ail/Lom bacterial outer membrane proteins: E. coli OmpX. • NMR in membranes • Barrel tilt • Structure determination Vogt & Schulz (1999) Structure 7:1301 Fernandez et al. (2004) J. Mol. Bio. 336:1211 SDS 14-O-PC SDS 14-O-PC Mahalakshmi et al. (2007) BBA Biomembranes, 1768:3216

  6. E. coli OmpX in magnetically oriented phospholipid bicelles. H2O D2O Mahalakshmi et al. (2007) BBA Biomembranes, 1768:3216

  7. E. coli OmpX in magnetically oriented phospholipid bicelles.

  8. assign refine back-calculate Mahalakshmi and Marassi (2008) Biochemistry, 47:6531

  9. Yersinia Pestis Ail outer membrane protein: production.

  10. Yersinia Pestis Ail outer membrane protein: refolding.

  11. Yersinia Pestis Ail outer membrane protein in micelles.

  12. Mycobacterium tuberculosis outer membrane protein OmpATb.

  13. Mycobacterium tuberculosis outer membrane protein OmpATb. • Search of FFAS Server yields homology to • BON (Bacterial OsmY and Nodulation) domain • 60 residues • αβ predicted fold • osmotic shock protection • interaction with membrane lipids • Yeats and Bateman (2003) TRENDS Biochem. Sci., 28:352

  14. Thank You OmpX R. Mahalakshmi (Burnham) Cecile Loudet (Burnham) Georg Schulz (A.L.U., Freiburg) Ail Candace Rypien (Burnham) Leigh Plesniak (U. San Diego) Greg Plano (U. Miami) OmpATb Peter Teriete (Burnham) Adrian Kolodzik (Burmham) Michael Niederweis (U. Alabama, Birmingham) TBBPC bicelles Erick Dufourc (U. Bordeaux) NIH/NIGMS Roadmap for Membrane Protein Structures

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