Localization of different epitopes in the structure of active PAI-1. (A) Cartoon representation of the crystal structure of PAI-1 in complex with the Fab fragment of MEDI-579 (PDB ID 6I8S) (132). The heavy and the light chain of the Fab fragment are colored cyan and green, respectively. The constant and variable heavy (CH1 and VH) and light (CL and VL) domains are indicated in the figure. Residues closer than 4 Å to MEDI-579 are indicated as spheres. Residues located in the RCL are colored red, residues residing in the exosite regions for the 37-loop and 60-loop of PAs are colored yellow and orange, respectively. (B) Localization of different epitopes of monoclonal antibodies (mAbs) as determined by mutagenesis studies. The epitopes of mAbs that prevent the interaction between PAI-1 and PAs are indicated as red (MA-42A2F6, MA-56A7C10, and MA-44E4) (235) and yellow (MA-124K1) (236) spheres. The epitopes of switching mAbs that bind to hF or the hF-s3A loop (MA-33H1F7, MA-55F4C12, and Mab2) (237, 238) are indicated as magenta spheres. The epitope of switching mAb MA-8H9D4 (239) that binds to the hI-s5A loop is indicated as green spheres. The epitopes of latency-inducing antibody MA-33B8 (240, 241), MA-H4B3 (90), and MA-159M12 (242) are indicated as cyan, orange, and brown spheres, respectively. (C) Localization of different epitopes of nanobodies as determined by mutagenesis studies (35). The epitope of substrate-inducing nanobody Nb98 is indicated by green and magenta spheres, whereas only the magenta spheres indicate the epitope of Nb64. The epitope of Nb93 that interferes with PAI-1/PA complex formation is indicated as red spheres. (D) Cartoon representation of the superimposed crystal structures of PAI-1 in complex with Nb42 [PDB ID 6GWN, 6GWP, and 6GWQ (52)], Nb64 [PDB ID 6GWN and 6GWP (52)], and Nb93 [PDB ID 6ZRV (41)]. Nb42, Nb64, and Nb93 are colored yellow, green, and magenta, respectively. Residues closer than 4 Å to Nb42 are indicated as yellow spheres. Residues closer than 4 Å to Nb64 are indicated as green spheres. Residues closer than 4 Å to Nb93 are indicated as red and magenta spheres. Red spheres represent residues located in the RCL of PAI-1, whereas magenta spheres represent residues located in the exosite binding regions for PAs.