Fig. S4.
Sequence conservation of interface residues in GLD-2 and GLD-3. (A) Circular dichroism (CD) spectra obtained for 0.15 mg/mL GLD-31–110 (blue line) and 0.18 mg/mL GLD-388–460 (red line) in 20 mM Tris pH 7.5, 150 mM NaCl, and 10% glycerol at 20 °C from 250 to 190 nm by using a 1-mm cuvette. For GLD-31–110, the strong negative peak at 202 nm indicate the presence of a random coil structure, whereas for GLD-388–460, the spectra indicate the presence of a globular structure. (B) Structure-based sequence alignment of GLD-3NT from C. elegans, Caenorhabditis brenneri, Caenorhabditis briggsae, and Caenorhabditis remanei. The secondary structure elements of C.e. GLD-3NT are shown above the sequence and colored according to Fig. 1A. Strictly and well-conserved residues are highlighted in dark and light purple, respectively. GLD-2-interacting residues are indicated with blue or pink dots, for highlighting interaction with the catalytic or central domain of GLD-2, respectively. Residues targeted for mutagenesis are shown with the substituted amino acid in black, above the sequences. (C, Left) Structure-based sequence alignment of GLD-2 from C. elegans (C.e.), H. sapiens (H.s.), D. melanogaster (D.m.), and X. laevis (X.l.) adapted from Fig. S3. The secondary structure elements of C.e. GLD-2 are shown above the sequence and colored according to Fig. 1A. Strictly and well conserved residues are highlighted in dark and light purple, respectively. Low conserved residues are highlighted in light pink. GLD-3–interacting residues are indicated with green dots. Active-site residues discussed in the text are indicated with an asterisk. (Right) Cartoon representation of the GLD-2–GLD-3 heterodimer in the same orientation as in Fig. 2B. Conserved interface residues in GLD-2 highlighted in the sequence alignment are annotated and shown as sticks.