Identification of the histidine-rich glycoprotein domains responsible for contact pathway inhibition

J Thromb Haemost. 2022 Apr;20(4):821-832. doi: 10.1111/jth.15631. Epub 2022 Jan 14.


Background: Previously, we showed that histidine-rich glycoprotein (HRG) binds factor (F) XIIa with high affinity, inhibits FXII autoactivation and FXIIa-mediated activation of FXI, and attenuates ferric chloride-induced arterial thrombosis in mice. Therefore, HRG downregulates the contact pathway in vitro and in vivo.

Objective: To identify the domains on HRG responsible for contact pathway inhibition.

Methods: Recombinant HRG domain constructs (N-terminal [N1, N2, and N1N2], proline-rich regions, histidine-rich region [HRR], and C-terminal) were expressed and purified. The affinities of plasma-derived HRG, HRG domain constructs, and synthetic HRR peptides for FXII, FXIIa, β-FXIIa, and polyphosphate (polyP) were determined using surface plasmon resonance, and their effects on polyP-induced FXII autoactivation, FXIIa-mediated activation of FXI and prekallikrein, the activated partial thromboplastin time (APTT), and thrombin generation were examined.

Results: HRG and HRG domain constructs bind FXIIa, but not FXII or β-FXII. HRR, N1, and N1N2 bind FXIIa with affinities comparable with that of HRG, whereas the remaining domains bind with lower affinity. Synthetic HRR peptides bind FXIIa and polyP with high affinity. HRG and HRR significantly inhibit FXII autoactivation and prolong the APTT. Like HRG, synthetic HRR peptides inhibit FXII autoactivation, attenuate FXIIa-mediated activation of prekallikrein and FXI, prolong the APTT, and attenuate thrombin generation.

Conclusion: The interaction of HRG with FXIIa and polyP is predominantly mediated by the HRR domain. Like intact HRG, HRR downregulates the contact pathway and contributes to HRG-mediated down regulation of coagulation.

Keywords: coagulation; factor XII; histidine-rich glycoprotein; polyphosphate; thrombosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Factor XII / metabolism
  • Factor XIIa / metabolism
  • Humans
  • Mice
  • Peptides / pharmacology
  • Polyphosphates
  • Prekallikrein* / metabolism
  • Proteins
  • Thrombin* / metabolism


  • Peptides
  • Polyphosphates
  • Proteins
  • histidine-rich proteins
  • Factor XII
  • Prekallikrein
  • Factor XIIa
  • Thrombin