Coagulation factor XII protease domain crystal structure

Pathak, Monika; Wilmann, P.G.; Awford, James; Li, C.; Hamad, B; Fischer, Peter M.; Dreveny, Ingrid; Dekker, Lodewijk V.; Emsley, Jonas

doi:10.1111/jth.12849

Cited by 48 publications

(47 citation statements)

References 34 publications

(48 reference statements)

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“…Recently, the crystal structure of the protease domain of human FXII in the zymogen‐like conformation was solved by using a Drosophila S2 cell–expressed construct. The structure identifies several interesting features, including areas of anionic charges around the active site cleft, which may play a key role in binding to substrates and potential inhibitors . While there are likely differences in glycosylation patterns and other post‐translational modifications in mammalian cells, the FXIIa light chain crystal structure provides valuable information for understanding FXIIa interactions with ligands.…”

Section: Fxii Biochemistry and Activationmentioning

confidence: 99%

Contact system revisited: an interface between inflammation, coagulation, and innate immunity

Long

Kenne

Jung

et al. 2016

Journal of Thrombosis and Haemostasis

267

286

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Summary. The contact system is a plasma protease cascade initiated by factor XII (FXII) that activates the proinflammatory kallikrein-kinin system and the procoagulant intrinsic coagulation pathway. Anionic surfaces induce FXII zymogen activation to form proteolytically active FXIIa. Bacterial surfaces also have the ability to activate contact system proteins, indicating an important role for host defense using the cooperation of the inflammatory and coagulation pathways. Recent research has shown that inorganic polyphosphate found in platelets activates FXII in vivo and can induce coagulation in pathological thrombus formation. Experimental studies have shown that interference with FXII provides thromboprotection without a therapy-associated increase in bleeding, renewing interest in the FXIIa-driven intrinsic pathway of coagulation as a therapeutic target. This review summarizes how the contact system acts as the cross-road of inflammation, coagulation, and innate immunity.

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Section: Fxii Biochemistry and Activationmentioning

confidence: 99%

Contact system revisited: an interface between inflammation, coagulation, and innate immunity

Long

Kenne

Jung

et al. 2016

Journal of Thrombosis and Haemostasis

267

286

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“…Interestingly, β bulges would accommodate structural changes without totally disrupting the β sheet . Further, the side chain of Leu251 contributes to the hydrophobic character of a pocket positioned directly in front of the S1 pocket, of special interest as coagulation protease substrates and inhibitors commonly bind this region . Substitution by proline is expected to alter protein native structure, potentially interfering but not suppressing protein folding, secretion and activity by preserving the hydrophobic nature of the highly conserved region.…”

Section: Discussionmentioning

confidence: 99%

Missense changes in the catalytic domain of coagulation factor X account for minimal function preventing a perinatal lethal condition

et al. 2019

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Introduction Inherited deficiencies in the coagulation pathway provide diversified models to investigate the molecular bases of perinatal lethality associated with null‐like variants. Differently from X‐linked haemophilias, homozygous/doubly heterozygous null variants in the rare autosomally inherited deficiency of factor X (FX) might be incompatible with perinatal survival. Aim To provide experimental evidence about the null/close‐to‐null FX function. Methods The residual secreted (ELISA) and functional (thrombin generation assays) protein levels associated with the novel nonsense (c.1382G>A; p.Trp461Ter) and missense (c.752T>C; p.Leu251Pro) variants, found in the proposita with life‐threatening symptoms at birth, were characterized through recombinant (r)FX expression. Results The rFX‐461Ter showed very low secretion and undetectable function. Expression and function of the predicted readthrough‐deriving missense variants (rFX‐461Tyr, rFX‐461Gln) were also severely impaired. These unfavourable features, due to nucleotide and protein sequence constraints, precluded functional readthrough over the 461 stop codon. Differently, the poorly secreted rFX‐251Pro variant displayed residual function that was characterized by anti‐TFPI aptamer‐based amplification or selective inhibition of activated FX function by fondaparinux in plasma and found to be reduced by approximately three orders of magnitude. Similarly to the rFX‐251Pro, a group of catalytic domain missense variants cause poorly secreted molecules with modest function in FX‐deficient patients with life‐threatening symptoms. Conclusions Our data, contributing to the knowledge of the very severe FX deficiency forms, support life‐saving requirement of trace FX function, clearly exemplified by the dysfunctional but not completely inactive rFX‐251Pro variant that, albeit with severely reduced function, is compatible with a residual activity ensuring minimal haemostasis and permitting perinatal survival.

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“…We have previously reported two zymogen-like crystal structures of the FXII protease spanning the protease domain alone (amino acids 354-596; Pathak et al, 2015). These are termed (i) FXIIc (PDB entry 4xde), which corresponds to a construct in which the N-terminus is blocked by the addition of two extra amino acids Arg-Ser and has a very low enzymatic activity, and (ii) FXIIac (PDB entry 4xe4; Pathak et al, 2015), which has the native N-terminal Val354 residue and a tenfold higher enzyme activity compared with FXIIc but a 1000-fold lower activity compared with FXIIa. FXIIac lacks the additional N-terminal residues of the HCR that are present in FXIIa, which we have speculated to be required for efficient catalytic activity (Pathak et al, 2015).…”

Section: Comparison Of Bfxiia His With Fxii Zymogen Protease Structuresmentioning

confidence: 99%

“…1). A further cleavage of FXIIa between residues Arg334 and Asn335 produces -factor XII (FXIIa), which includes the light chain and nine residues of the heavy chain, which are termed the heavy-chain remnant (HCR; Pathak et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Crystal structures of the recombinant β-factor XIIa protease with bound Thr-Arg and Pro-Arg substrate mimetics

Pathak

Manna

et al. 2019

Acta Cryst Sect D Struct Biol

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Coagulation factor XII (FXII) is a key initiator of the contact pathway, which contributes to inflammatory pathways. FXII circulates as a zymogen, which when auto-activated forms factor XIIa (FXIIa). Here, the production of the recombinant FXIIa protease domain (FXIIa His ) with yields of $1-2 mg per litre of insect-cell culture is reported. A second construct utilized an N-terminal maltose-binding protein (MBP) fusion (MBP-FXIIa His ). Crystal structures were determined of MBP-FXIIa His in complex with the inhibitor d-Phe-Pro-Arg chloromethyl ketone (PPACK) and of FXIIa His in isolation. The FXIIa His structure revealed that the S2 and S1 pockets were occupied by Thr and Arg residues, respectively, from an adjacent molecule in the crystal. The Thr-Arg sequence mimics the P2-P1 FXIIa cleavage-site residues present in the natural substrates prekallikrein and FXII, and Pro-Arg (from PPACK) mimics the factor XI cleavage site. A comparison of the FXIIa His structure with the available crystal structure of the zymogen-like FXII protease revealed large conformational changes centred around the S1 pocket and an alternate conformation for the 99-loop, Tyr99 and the S2 pocket. Further comparison with activated protease structures of factors IXa and Xa, which also have the Tyr99 residue, reveals that a more open form of the S2 pocket only occurs in the presence of a substrate mimetic. The FXIIa inhibitors EcTI and infestin-4 have Pro-Arg and Phe-Arg P2-P1 sequences, respectively, and the interactions that these inhibitors make with FXIIa are also described. These structural studies of FXIIa provide insight into substrate and inhibitor recognition and establish a scaffold for the structure-guided drug design of novel antithrombotic and antiinflammatory agents.

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Coagulation factor XII protease domain crystal structure

Cited by 48 publications

References 34 publications

Contact system revisited: an interface between inflammation, coagulation, and innate immunity

Contact system revisited: an interface between inflammation, coagulation, and innate immunity

Missense changes in the catalytic domain of coagulation factor X account for minimal function preventing a perinatal lethal condition

Crystal structures of the recombinant β-factor XIIa protease with bound Thr-Arg and Pro-Arg substrate mimetics

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