Bone morphogenetic proteins (BMPs) 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 (BMPRs) of type I and type II, and a promiscuity has been reported. In this study, we used biolayer interferometry to perform parallel real-time biosensing and to deduce the kinetic parameters (ka, kd) and the equilibrium constant (KD) for a large range of BMP/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 the type-II BMPR receptor 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 in a ternary complex 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.

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See supplementary material at https://www.scitation.org/doi/suppl/10.1116/6.0000926 for S1 TABLE 1. Detailed literature study table summarizing the KD (nM) of the BMP/type-I and type-II BMPR interaction couples; S2 FIGURE 1. Image of a gel electrophoresis showing all the used BMPs, ALKs and type-II BMPR; S3 TABLE 2. Detailed kinetic tables indicating the KD (nM), ka (M-1.s-1), kd (s-1) and R2 of the BMP/type-I interaction couples; S4 TABLE 3. Detailed kinetic tables indicating the KD (nM), ka (M-1.s-1), kd (s-1) and R2 of the BMP/type-II interaction couples; S5 FIGURE 2. Binding kinetics between non-glycosylated and glycosylated BMP-4 with ALK3; S6 FIGURE 3. SPR binding curve for BMP-2/ALK3; S7 FIGURE 4. BLI binding kinetics of BMP-2 with ALK2 and ACTR-IIB in two confirmations using 1:1 fit.

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