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. 2014 Feb 17;204(4):477–486. doi: 10.1083/jcb.201311014

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© 2014 Richter et al.

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Figure 2. — Analysis of MiD51 binding to GDP and ADP. (A) Raw heats of binding obtained using ITC by titration of MiD51^ΔN118 with nucleotide. (B) Table of binding constants obtained by fitting ITC data to a single-state binding model. NB denotes no binding. Asterisk denotes n = 2. (C) Nucleotide binding pocket of MiD51 harboring GDP. Key MiD51^ΔN118 residues are shown as sticks and GDP is shown in cyan. Hydrogen bonds are indicated by black dotted lines. (D) Nucleotide binding pocket of MiD51 harboring ADP. Key MiD51^ΔN118 residues are shown as sticks and ADP is shown in magenta. Hydrogen bonds are indicated by black dotted lines. (E) Superposition of GDP-MiD51 (cyan) and ADP-MiD51 (magenta). (F) Summary of MiD51 mutants evaluated in immunofluorescence assays for Drp1 recruitment. (G) MiD51^TALEN MEFs were induced to express MiD51-GFP or MiD51^NBD-GFP with 4-OHT and mitochondrial morphology counted. Only MEFs with low-level GFP fluorescence were scored. n = 3; mean ± SEM; 100 cells counted/condition.