1. Volume 23, Issue 6, Pages 1116-1122 (June 2015)
NMR Structure of Francisella tularensis Virulence Determinant Reveals Structural Homology to Bet v1 Allergen Proteins 
James Zook, Gina Mo, Nicholas J. Sisco, Felicia M. Craciunescu, Debra T. Hansen, Bobby Baravati, Brian R. Cherry, Kathryn Sykes, Rebekka Wachter, Wade D. Van Horn, Petra Fromme 
Structure 
Volume 23, Issue 6, Pages (June 2015)
DOI: /j.str
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2. Structure 2015 23, 1116-1122DOI: (10.1016/j.str.2015.03.025)
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3. Figure 1 Assigned 2D 15N-HSQC Spectrum of Flpp3sol
The NMR spectrum displays a wide proton dispersion with narrow peak widths, indicative of a protein with significant β-sheet and α-helix content.
Structure  , DOI: ( /j.str )
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4. Figure 2
Structure of Flpp3sol and Model of Flpp3 Inserted into the Membrane
(A) Overlay of the 20 lowest-energy models after water bath refinement.
(B) Cartoon model of the lowest-energy model of Flpp3sol. Residues F37, S41, A61, D63, and S66 may be dynamically important as per initial relaxation results, and are shown in violet.
(C and D) The transmembrane helix (residues Met1 to Thr25) modeled onto the N terminus of Flpp3sol and inserted into a POPC/POPE membrane. MD production simulations were run for 1 ns with experimental restraints followed by another 1 ns without restraints. The hydrophobic region of the helix spans the entire length of the membrane with a positively charged N terminus (not seen). Model is rotated 180° to show both sides of the protein. An electrostatic surface is plotted onto the model to show polarity between the membrane proximal region (blue/positive) and the membrane distal region (red/negative).
Structure  , DOI: ( /j.str )
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5. Figure 3
Comparison of Flpp3sol and Structurally Homologous Bet v1 Protein Birch Pollen Allergen
(A and B) Secondary structure and overall fold topology of Flpp3sol (A) and structurally homologous Bet v1 protein birch pollen allergen (1FM4) (B). The Bet v1 proteins are larger due to two extra β strands that distort the first α helix (purple).
(C and D) Electrostatic surface plotted onto Flpp3sol (C) and 1FM4 (D). Flpp3sol has a polarized surface with a negatively charged (red) side and a positively charged (blue) region compared with 1FM4.
(E and F) The internal cavity is shown for Flpp3sol (E) and 1FM4 (F). The two extra β strands (purple) in 1FM4 provide additional cavity space that is limited by Flpp3sol.
Structure  , DOI: ( /j.str )
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6. Figure 4 15N-NMR Relaxation Studies
R1 data do not vary considerably between residues and demonstrates an overall constant rate of 2.2 ± 0.2 s−1. R2 data reveal a fairly constant relaxation rate throughout the structured regions of Flpp3sol. Flexible termini show slightly decreased values while several residues in the loop region reveal much greater R2 relaxation rates, which may be the result of chemical exchange. In the steady-state heteronuclear NOE experiment, NOE values are relatively large and unchanging. The histidine tail did show a negative NOE value but was left out of the analysis because of inability to resolve individual histidine residues. Error bars reflect ±1 SD from the calculated mean.
Structure  , DOI: ( /j.str )
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