Factor XII and kininogen asymmetric assembly with gC1qR/C1QBP/P32 is governed by allostery

Kaira, Bubacarr G.; Slater, Alexandre; McCrae, Keith R.; Dreveny, Ingrid; Sumya, Um-May; Mutch, Nicola J.; Searle, Mark; Emsley, Jonas

Abstract

The contact system is composed of Factor XII (FXII), prekallikrein (PK) and co-factor kininogen (HK). The globular C1q receptor (gC1qR) has been shown to interact with FXII and HK. We reveal the FXII fibronectin type II domain (FnII) binds gC1qR in a Zn2+ dependent fashion and determined the complex crystal structure. FXIIFnII binds the gC1qR trimer in an asymmetric fashion with residues Arg36 and Arg65 forming contacts with two distinct negatively charged pockets. gC1qR residues Asp185 and His187 coordinate a Zn2+ adjacent to the FXII binding site and a comparison with the ligand free gC1qR crystal structure reveals the anionic G1-loop becomes ordered upon FXIIFnII binding. Additional conformational changes in the region of the Zn2+ binding site reveal an allosteric basis for Zn2+ modulation of FXII binding. Mutagenesis coupled with SPR demonstrate the gC1qR Zn2+ site contributes to FXII binding and plasma based assays reveal gC1qR stimulates coagulation in a FXII-dependent manner. Analysis of the binding of HK domain 5 (HKD5) to gC1qR shows only one high affinity binding site per trimer. Mutagenesis studies identify a critical G3-loop located at the center of the gC1qR trimer suggesting steric occlusion as the mechanism for HKD5 asymmetric binding. Gel filtration experiments reveal that gC1qR clusters FXII and HK into a higher order 500kDa ternary complex. These results support the conclusion that extracellular gC1qR can act as a chaperone to cluster contact factors which may be a prelude for initiating the cascades which drive bradykinin generation and the intrinsic pathway of coagulation.

Citation

Kaira, B. G., Slater, A., McCrae, K. R., Dreveny, I., Sumya, U., Mutch, N. J., …Emsley, J. (2020). Factor XII and kininogen asymmetric assembly with gC1qR/C1QBP/P32 is governed by allostery. Blood, 136(14), 1685–1697. https://doi.org/10.1182/blood.2020004818

• Journal Article Type: Article
• Acceptance Date: May 13, 2020
• Online Publication Date: Jun 19, 2020
• Publication Date: Oct 1, 2020
• Deposit Date: Jul 15, 2020
• Publicly Available Date: Jun 20, 2021
• Journal: Blood
• Print ISSN: 0006-4971
• Electronic ISSN: 1528-0020
• Publisher: American Society of Hematology
• Peer Reviewed: Peer Reviewed
• Volume: 136
• Issue: 14
• Pages: 1685–1697
• DOI: https://doi.org/10.1182/blood.2020004818
• Keywords: Immunology; Cell Biology; Biochemistry; Hematology
• Public URL: https://nottingham-repository.worktribe.com/output/4693050
• Publisher URL: https://ashpublications.org/blood/article-abstract/doi/10.1182/blood.2020004818/461064/Factor-XII-and-kininogen-asymmetric-assembly-with?redirectedFrom=fulltext
• Additional Information: This research was originally published in Blood. Bubacarr G Kaira,; Alexandre Slater, Keith R McCrae, Ingrid Dreveny, Um-May Sumya, Nicola J Mutch,Mark Searle, Jonas Emsley. Factor XII and kininogen asymmetric assembly with gC1qR/C1QBP/P32 is governed by allostery . Blood. Year; © the American Society of Hematology.