human bid bh3 peptide (h-sesqediirniarhlaqvgdsmdrsippglvnglnh2)  (GenScript corporation)

Journal: eLife

Article Title: Contingency and chance erase necessity in the experimental evolution of ancestral proteins

doi: 10.7554/eLife.67336

Figure Lengend Snippet: ( A ) Protein binding specificities of extant BCL-2 family members. Human MCL-1 (hsMCL-1, purple) strongly binds BID (blue) and NOXA (red), while human BCL-2 (hsBCL-2, green) strongly binds BID but not NOXA. ( B ) Crystal structures of MCL-1 (purple) bound to NOXA (red, PDB 2nla), and BCL-xL (green, a closely related paralog of BCL-2) bound to BID (blue, PDB 4qve). ( C ) Reduced maximum likelihood phylogeny of BCL-2 family proteins. Purple bar, MCL-1 class; green bar, BCL-2 class. The phylogeny was rooted using as outgroups the paralogs BOX, BAK, and BAX (black bar). Heatmaps indicate BID (blue) and NOXA (red) binding measured using the luciferase assay. Each shaded box shows the normalized mean of three biological replicates. Red dotted lines, interval during which NOXA binding was lost, yielding BID specificity in the BCL-2 proteins of vertebrates (green box). Purple box, vertebrate MCL-1. Silhouettes, representative species in each terminal group. AncMB1-M and -B are alternative reconstructions using different approaches to alignment ambiguity (see Materials and methods). For complete phylogeny, see .

Article Snippet: Peptide, recombinant protein , BID , GenScript , This Study , Human BID peptide used for fluorescence polarization (see Materials and methods).

Techniques: Protein Binding, Binding Assay, Luciferase

Journal: eLife

Article Title: Contingency and chance erase necessity in the experimental evolution of ancestral proteins

doi: 10.7554/eLife.67336

Figure Lengend Snippet: ( A ) BID fluorescence polarization for hsMCL-1 variants evolved to lose NOXA binding. Bars are the mean of three replicates; error bars, SD. mFP, normalized measured fluorescent polarization. Kd estimates are shown below in the table. ( B ) Same as ( A ), but for NOXA binding. ( C ) BID fluorescence polarization for hsBCL-2 variants evolved to gain NOXA binding. ( D ) Same as ( C ), but for NOXA binding.

Article Snippet: Peptide, recombinant protein , BID , GenScript , This Study , Human BID peptide used for fluorescence polarization (see Materials and methods).

Techniques: Fluorescence, Binding Assay

Journal: eLife

Article Title: Contingency and chance erase necessity in the experimental evolution of ancestral proteins

doi: 10.7554/eLife.67336

Figure Lengend Snippet:

Article Snippet: Peptide, recombinant protein , BID , GenScript , This Study , Human BID peptide used for fluorescence polarization (see Materials and methods).

Techniques: Recombinant, Fluorescence, Software

Journal: bioRxiv

Article Title: Conformational states of the cytoprotective protein Bcl-xL

doi: 10.1101/2020.03.25.007740

Figure Lengend Snippet: (A) Representative calorimetric data (top) and integrated heat (bottom) are shown as functions of BID BH3 /Bcl-xL molar ratio. The data were corrected for non-specific binding by subtracting control ITC titrations performed by titrating peptide into buffer or empty nanodiscs without Bcl-xL. Solid lines (red) are the best fits of the binding isotherms to a single-site binding model, used to extract the values of the dissociation constant (K d ). (B) Plot of the thermodynamic parameters for each calorimetric titration. Values represent the average of triplicate or duplicate titrations and erroro bars represent standard deviation.

Article Snippet: The BID BH3 peptide, corresponding to residues 80-99 in the BH3 domain of human BID, was obtained commercially (GenScript).

Techniques: Binding Assay, Control, Titration, Standard Deviation