Extended Data Fig. 1: Crystal structures of unphosphorylated PINK1.
a, Constructs used in crystal structures of TcPINK1 (PDB: 5OAT11 and 5YJ912). Phosphomimetics substituted for Ser205 (equivalent to Ser202 in PhPINK1) and other residues, although one structure (PDB: 5OAT) still showed phosphorylation at Ser207 (equivalent to Ser204 in PhPINK1, and Ser230 in HsPINK1, see below). Constructs were further engineered as indicated and described11,12. b, Depiction of TcPINK1 crystal structures, with key features indicated. c, 2|Fo|–|Fc| electron density contoured at 1.5σ, covering the PhPINK1(D334A) molecule in the asymmetric unit. Right, detail for regions of interest including the αC helix and Ser202-containing loop and the N-helix. d, Our crystal structure of unphosphorylated PhPINK1(D334A) (compare Fig. 1) in the same orientation as in b, for comparison. e, Two different zoomed views of the ATP-binding site of unphosphorylated PhPINK1, with an ATP molecule modelled in from PDB 2PHK (ref. 42) as before10. Left, highlighting key features common to most active kinases, including the P-loop (grey), αC helix (orange) and activation segment (pink). The DFG and HRD motifs (dotted outlines) which include Asp357 involved in coordination of Mg2+, and the substrate-binding Asp334 involved in phosphoryl transfer (and mutated to Ala in the crystal structure) are indicated. Glu214 and Lys193 form a crucial salt bridge to coordinate ATP. Right, assembled catalytic (C) and regulatory (R) spines24 in red and blue, respectively, indicative of an active kinase conformation. f, Crystal contact involving N-helix residue Trp129, which binds to the N-lobe of a symmetry related molecule. ASU, asymmetric unit.
