The activation peptide of coagulation factor IX and X serves as a high affinity receptor to cationic ligands

Griessner, A.; Zögg, Thomas; Brandstetter, Hans

doi:10.1160/th13-01-0051

Cited by 3 publications

(2 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Inhibition of furin by Nb14 can conveniently be monitored in limited proteolysis experiments with the furin substrate coagulation factor X 31 . Maturation of it requires cleavage by furin in between the catalytic domain and the EGF-2 domain.…”

Section: Resultsmentioning

confidence: 99%

The structure of a furin-antibody complex explains non-competitive inhibition by steric exclusion of substrate conformers

Dahms

Creemers

Schaub

et al. 2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

Proprotein Convertases (PCs) represent highly selective serine proteases that activate their substrates upon proteolytic cleavage. Their inhibition is a promising strategy for the treatment of cancer and infectious diseases. Inhibitory camelid antibodies were developed, targeting the prototypical PC furin. Kinetic analyses of them revealed an enigmatic non-competitive mechanism, affecting the inhibition of large proprotein-like but not small peptidic substrates. Here we present the crystal structures of furin in complex with the antibody Nb14 and of free Nb14 at resolutions of 2.0 Å and 2.3 Å, respectively. Nb14 binds at a site distant to the substrate binding pocket to the P-domain of furin. Interestingly, no major conformational changes were observed upon complex formation, neither for the protease nor for the antibody. Inhibition of furin by Nb14 is instead explained by steric exclusion of specific substrate conformers, explaining why Nb14 inhibits the processing of bulky protein substrates but not of small peptide substrates. This mode of action was further supported by modelling studies with the ternary factor X-furin-antibody complex and a mutation that disrupted the interaction interface between furin and the antibody. The observed binding mode of Nb14 suggests a novel approach for the development of highly specific antibody-based proprotein convertase inhibitors.

show abstract

Section: Resultsmentioning

confidence: 99%

The structure of a furin-antibody complex explains non-competitive inhibition by steric exclusion of substrate conformers

Dahms

Creemers

Schaub

et al. 2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Light and heavy chains remain covalently linked via a disulfide bond. In both factor IX and factor X, the activation peptide harbours an intriguing high‐affinity binding site for benzamidine/arginine, exceeding the affinity towards the active site by an order of magnitude . By contrast, macromolecular factor IX/X inhibitors such as serpins or Kunitz inhibitors such as BPTI do not bind to the activation peptide but to the active site exclusively.…”

mentioning

confidence: 99%

Maturation of coagulation factor IX during Xase formation as deduced using factor VIII‐derived peptides

2019

Self Cite

View full text Add to dashboard Cite

Blood coagulation involves extrinsic and intrinsic pathways, which merge at the activation step of blood coagulation factor X to factor Xa. This step is catalysed by the extrinsic or intrinsic Xase, which consists of a complex of factor VIIa and its cofactor tissue factor or factor IXa (FIXa) and its cofactor coagulation factor VIIIa (FVIIIa). Upon complex formation with FVIIIa, FIXa is conformationally activated to the Xase complex. However, the mechanistic understanding of this molecular recognition is limited. Here, we examined FVIIIa‐FIXa binding in the context of FIXa's activation status. Given the complexity and the labile nature of FVIIIa, we decided to employ two FVIII‐derived peptides (558‐loop, a2 peptide) to model the cofactor binding of FIX(a) using biosensor chip technology. These two FVIII peptides are known to mediate the key interactions between FVIIIa and FIXa. We found both of these cofactor mimetics as well as full‐length FVIIIa bind more tightly to zymogenic FIX than to proteolytically activated FIXa. Consequently and surprisingly, we observed that the catalytically inactive FIX zymogen can outcompete the activated FIXa from the complex with FVIIIa, resulting in an inactive, zymogenic Xase complex. By contrast, the thrombophilic Padua mutant FIXa‐R170 in complex with the protein–substrate analogue BPTI bound tighter to FVIIIa than to the zymogen form FIX‐R170L, suggesting that the active Xase complex preferentially forms in the Padua variant. Together, these results provide a mechanistic basis for the thrombophilic nature of the FIX‐R170L mutant and suggest the existence of a newly discovered safety measure within the coagulation cascade.

show abstract

Sirtilins – the new old members of the vitamin K‐dependent coagulation factor family

Dahms

Demir

Huesgen

et al. 2019

Journal of Thrombosis and Haemostasis

View full text Add to dashboard Cite

Essentials Blood coagulation is driven by vitamin K (VK)‐dependent proteases.We have identified and characterized ‘sirtilin’ as an additional VK‐dependent protease.Sirtilins emerged early in the evolution of the coagulation system of vertebrates.Ubiquitous occurrence might indicate an important functional role of sirtilins. SummaryBackgroundVitamin K (VK)‐dependent proteases are major players in blood coagulation, including both the initiation and the regulation of the cascade. Five different members of this protease family have been described, comprising the following coagulation factors: factor VII, FIX, FX, protein C (PC), and prothrombin (FII). FVII, FIX, FX and PC share a typical domain architecture, with an N‐terminal γ‐carboxyglutamate (Gla) domain, two epidermal growth factor‐like (EGF) domains, and a C‐terminal trypsin‐like serine protease (SP) domain.ObjectivesWe have identified uncharacterized proteins in snake genomes showing the typical Gla–EGF1–EGF2–SP domain architecture but relatively low sequence conservation compared to known VK‐dependent proteases. On the basis of sequence analysis, we hypothesized that these proteins are functional members of the VK‐dependent protease family.Methods/resultsUsing phylogenetic analyses, we confirmed the so‐called ‘sirtilins’ as an additional VK‐dependent protease class. These proteases were found in several vertebrates, including jawless fish, cartilaginous fish, bony fish, reptiles, birds, and marsupials, but not in other mammals. The recombinant zymogen form of Thamnophis sirtalis sirtilin was produced by in vitro renaturation, and was activated with human activated FXI. The activated form of sirtilin proteolytically cleaved peptide and protein substrates, including prothrombin. Mass spectrometry‐based substrate profiling of sirtilin revealed a narrower sequence specificity than those of FIX and FX.ConclusionsThe ubiquitous occurrence of sirtilins in many vertebrate classes might indicate an important functional role. Understanding the detailed functions of sirtilins might contribute to a deeper understanding of the evolution and function of the vertebrate coagulation system.

show abstract

The activation peptide of coagulation factor IX and X serves as a high affinity receptor to cationic ligands

Abstract: Moix N, et al. Position of nonmuscle myosin heavy chain IIA (NMMHC-IIA) mutations predicts the natural history of MYH9-related disease.

Cited by 3 publications

References 14 publications

The structure of a furin-antibody complex explains non-competitive inhibition by steric exclusion of substrate conformers

The structure of a furin-antibody complex explains non-competitive inhibition by steric exclusion of substrate conformers

Maturation of coagulation factor IX during Xase formation as deduced using factor VIII‐derived peptides

Sirtilins – the new old members of the vitamin K‐dependent coagulation factor family

Contact Info

Product

Resources

About