Figure 1.

In Vitro Characterization of OGT_L254F

(A) Schematic representation of OGT highlighting the intellectual disability-associated mutations and all the constructs used in this study.

(B) Scatterplot showing OGT activity against deglycosylated HEK-293 cell lysate, with the data averaged from six replicates and the error bars showing SD. See also Figure S1B.

(C) Superposition of the TPR_WT/L254F crystal structures at the site of mutation. The gray and colored cartoons are that of TPR_WT (PDB: 1W3B; Jínek et al., 2004) and TPR_L254F (PDB: 6EOU) structures, respectively.

(D) Overlay of the chimeric OGT_WT/L254F structures. The wild-type structure is colored gray, while the mutant structure is colored to reflect the positional shift of each C_α atom between the two structures.

(E) Graph showing the positional shift between equivalent C_α atoms between chimeric OGT_WT (PDB: 4XIF [Pathak et al., 2015] and PDB: 1W3B [Jínek et al., 2004]) and OGT_L254F (PDB: 4XIF [Pathak et al., 2015] and PDB: 6EOU) as a function of residue number.

L, L254F; OGT, O-GlcNAc transferase; sTPR, simplified TPR; TPR, tetratricopeptide repeat; W, wild-type.