Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Crystal structure of human plasma β-FXIIa. (A) Stereo ribbon diagram (cyan) of β-FXIIa in complex with covalent compound 1 inhibitor in “standard” orientation. Compound 1 is shown in green sticks. The side chains of key β-FXIIa residues are shown as magenta sticks: His57, Asp102, and Ser195 of the catalytic triads, Asp189 at the base of the S1 specificity pocket. The side chains of N-glycosylated Asn74 and NacGlc-disaccharide are shown in cyan sticks. Some surface loops discussed in the text are labeled and shown in pink. The heavy chain remnant of β-FXIIa is shown in red. Hydrogen bonds are indicated by black dotted lines. (B) The structure of β-FXIIa in complex with noncovalent benzamidine inhibitor is represented as in panel A. Six disulfide bonds are indicated and are shown in yellow. (C) Superposition of the β-FXIIa (benzamidine [magenta] and β-FXIIa) compound 1 (cyan) crystal structures. The catalytic domains are superimposed by aligning the Cα atoms of residues 16-244 and are presented as cartoon diagrams. Benzamidine (orange), compound 1 (green), and residues of the catalytic triad are shown in a stick representation.

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques: Labeling

Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Surface charge of the β-FXIIa protease domain. (A) Solid surface colored according to the electrostatic potential calculated from the crystal structure of β-FXIIa in complex with compound 1. Positive surface is shown in blue and negative is in red. Some surface loops surrounding the active site cleft and positions of the substrate-binding sites (S4-S4′) are labeled. Noncovalent inhibitors, sugar residues, and sulfate ions are shown as sticks. (B) Thrombin surface charge was calculated from the crystal structure (PDB code 4UFD) of the enzyme in complex with the benzamidine-containing moiety. The inhibitor is shown in green sticks and the view of the thrombin molecule is that of the β-FXIIa molecule orientation shown in panel A. (C) Close-up stereo view of the FXIIa substrate from P3 to P4′ (ball and stick in cyan) interacting with the surface of the substrate-binding area. The residues P3 to P4′ modeled into the active site (see “Materials and methods”) according to the canonical conformation are Lys-Pro-Arg-Ile-Val-Gly-Gly from the FXI amino acid sequence (residues 385-391; the scissile peptide bond is between the underlined residues). The positions of several FXIIa catalytic domain residues, which can form hypothetical interactions with the substrate in the encounter complex are indicated.

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques: Binding Assay, Labeling, Sequencing

Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Sequences around the scissile peptide bonds of the β-FXIIa substrates FXI and kallikrein and of the β-FXIIa endogenous inhibitors serpin C1, antithrombin, and PAI-1

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques:

Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Two distinct intermolecular contacts between symmetry related molecules of β-FXIIa protease domain in the crystal packing. (A) First extensive interface. Ribbon representation of 2 protease domains shown in cyan and magenta. Contacting residues are labeled and shown as sticks. The residues of the neighboring molecule are denoted by the “′” mark. Salt bridges and hydrogen bonds are depicted by black dotted lines. Compound 1, buried in the active site, is shown as balls. Positions of the catalytic triad for both molecules and the heavy chain remnant for one molecule (on the right) are indicated. (B) Second contact region. The same as in panel A except the heavy chain remnant of the β-FXIIa molecule on the left (yellow cartoon) is shown in red.

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques: Labeling

Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Binding interactions of small molecule inhibitors to β-FXIIa. (A) Close-up view of the inhibitor binding area in the benzamidine complex (gray ribbon). The benzamidine molecule (green sticks) bound to the S1 site, the residues defining the interaction with the inhibitors, the residues of the catalytic triad, and the bound phosphate ions (yellow) are shown as sticks. The electron density maps (blue) around the bound inhibitor and the ion are countered at 1σ. (B) The compound 1–β-FXIIa complex. The same view as in panel A.

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques: Binding Assay

Journal: Blood Advances

Article Title: Structures of human plasma β–factor XIIa cocrystallized with potent inhibitors

doi: 10.1182/bloodadvances.2018016337

Figure Lengend Snippet: Structural comparison of the catalytic domains of β-FXIIa and FXIIc. Superposition of the β-FXIIa–compound 1 (cyan) reported in this study and the zymogen form of the FXII catalytic domain (FXIIc, residues 16-244; PDB code 4XDE, gray cartoon). The greatest conformational changes between 2 structures are indicated by dashed lines.

Article Snippet: Materials Human plasma β-FXIIa and PK were purchased from Molecular Innovations (Novi, MI).

Techniques: