High throughput measurements of BMP/BMP receptors interactions using bio-layer interferometry

Archive ouverte

Khodr, Valia | Machillot, Paul | Migliorini, Elisa | Reiser, Jean-Baptiste | Picart, Catherine

Edité par CCSD -

Bone morphogenetic proteins (BMP) are an important family of growth factors playing a role in a large number of physiological and pathological processes, including bone homeostasis, tissue regeneration and cancers. In vivo , BMPs bind successively to both BMP receptors (BMPR) of type I and type II, and a promiscuity has been reported. In this study, we used bio-layer interferometry to perform parallel real-time biosensing and to deduce the kinetic parameters (k a , k d ) and the equilibrium constant (K D ) for a large range of BMPs/BMPR combinations in similar experimental conditions. We selected four members of the BMP family (BMP-2, 4, 7, 9) known for their physiological relevance and studied their interactions with five type-I BMP receptors (ALK1, 2, 3, 5, 6) and three type-II BMP receptors (BMPR-II, ACTR-IIA, ACTR-IIB). We reveal that BMP-2 and BMP-4 behave differently, especially regarding their kinetic interactions and affinities with the type-II BMPR. We found that BMP-7 has a higher affinity for ACTR-IIA and a tenfold lower affinity with the type-I receptors. While BMP-9 has a high and similar affinity for all type-II receptors, it can interact with ALK5 and ALK2, in addition to ALK1. Interestingly, we also found that all BMPs can interact with ALK5. The interaction between BMPs and both type-I and type II receptors immobilized on the same surface did not reveal further cooperativity. Our work provides a synthetic view of the interactions of these BMPs with their receptors and paves the way for future studies on their cell-type and receptor specific signaling pathways. . Abbreviations BMP-2, bone morphogenetic protein 2; BMP-4, bone morphogenetic protein 4; BMP-7, bone morphogenetic protein 7; BMP-9, bone morphogenetic protein 9; BMPR, Bone morphogenetic protein receptor; ALK1, Activin receptor-like kinase 1; ALK2, Activin receptor-like kinase 2; ALK3, Activin receptor-like kinase 3; ALK5, Activin receptor-like kinase 5; ALK6, Activin receptor-like kinase 6; ECD, extracellular domain; TGFβ, transforming growth factor; TGFβR, transforming growth factor receptor; BLI, Bio-layer interferometry; SPR, Surface plasmon resonance.

• Description
• Sujet/Public
• Outil
• Outil d'anticipation
• Mémoire et thèse
• Gestion