1. Volume 19, Issue 10, Pages 1525-1534 (October 2011)
Structural Insight into the Mycobacterium tuberculosis Rv0020c Protein and Its Interaction with the PknB Kinase 
Christian Roumestand, Jade Leiba, Nathalie Galophe, Emmanuel Margeat, André Padilla, Yannick Bessin, Philippe Barthe, Virginie Molle, Martin Cohen-Gonsaud 
Structure 
Volume 19, Issue 10, Pages (October 2011)
DOI: /j.str
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2. Figure 1
Superimposition of the [1H,15N] HSQC Spectra of Full-Length, N-Terminal, and C-Terminal Domains of Rv0020c
Superimposition of the [1H,15N] HSQC spectra obtained at 700 MHz (pH 6.8 and 20°C), of Rv0020c full-length (black), Rv0020c_FHA (blue), and Rv0020c_Nter (red). The averaged chemical shift differences (Δδ) as a function of the protein sequence are given in Figure S1. See also Figure S2.
Structure  , DOI: ( /j.str )
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3. Figure 2 Cartoon Representation of the Rv0020c_FHA Structure
(A) The N-terminal part of the protein from Gly430 to Gly433 is disordered and represented in gray.
(B) Representation of the final ensemble of 30 NMR structures of Rv0020c_FHA.
Structure  , DOI: ( /j.str )
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4. Figure 3 Cartoon Representation of the Rv0020c_Nter Structure
(A) The N-terminal part of the protein from Met1 to Val32 is disordered and represented in gray.
(B) Representation of the final ensemble of 30 NMR structures of Rv0020c_Nter.
See also Figure S3.
Structure  , DOI: ( /j.str )
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5. Figure 4
In Vitro Phosphorylation of the Rv0020c Derivatives by PknB_331
Purified Rv0020c_WT and Rv0020c_T116A were incubated with PknB_331 and [γ-33]ATP. Samples were separated by SDS-PAGE, stained with Coomassie blue (upper panel) and visualized by autoradiography (lower panel) after overnight exposure to a film, as indicated. Upper bands reflect the autophosphorylation activity of Rv0020c and the lower bands correspond to the autophosphorylation signal of PknB_331. See also Figures S4 and S5.
Structure  , DOI: ( /j.str )
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6. Figure 5 Fluorescence Anisotropy Binding Profile
Fluorescence anisotropy binding profile for binding of Atto647N-labeled Rv0020c_FHA (4 nM) with PknB_331 (A), PknB_279 (B), PknB_331_ T294A (C), PknB_331_T309A (D). See also Figure S6.
Structure  , DOI: ( /j.str )
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7. Figure 6
Superimpositions on Selected of Rv0020c_FHA Alone and in the Presence of Peptides
Superimpositions of zooms on selected resonances in the full [1H,15N] HSQC spectra of uniformly labeled samples Rv0020c_FHA (50 μM) alone solution (black) and in the presence of 500 μM concentrations of Pep_294 (red) or Pep_309 (blue) (A), or in the presence of Pep_294/309 (500 μM) (pink) (B). Clearly, the bound Pep_294/309 peptide splits into two unequal populations that occupy two distinct sites at the protein surface. These two sites correspond to the binding sites already found either for Pep_294 (75%, as measured from the relative cross-peak intensity) or for Pep_309 (25%). Spectra were recorded at 500 MHz (20°C and pH 6.8). The corresponding full spectra are presented as Figures S8 and S9. See also Figure S7.
Structure  , DOI: ( /j.str )
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8. Figure 7
Amide Averaged Chemical Shift Variations (Δδ) as a Function of the Protein Sequence
Δδ have been measured between [1H,15N] HSQC spectra recorded at 500 MHz (20°C and pH 6.8) on 50 μM 15N-uniformly labeled samples of Rv0020c_FHA before and after addition of 500 μM concentrations of unlabeled peptides Pep_294 (A) or Pep_308 (C), with Δδ = [ΔδH2+(ΔδN.γN/γH)2]0.5. The dotted line stands for the standard deviation (0.13 ppm). The corresponding Van der Waals diagram of the Rv0020c_FHA NMR structure are presented in (B) and (D), with a color gradient indicative of the Δδ value: from gray (Δδ≈0) to red (Δδmax > 0.75 ppm).
Structure  , DOI: ( /j.str )
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