Bone Morphogenetic Protein Receptor Complexes on the Surface of Live Cells: A New Oligomerization Mode for Serine/Threonine Kinase Receptors

Abstract

The bone morphogenetic proteins (BMPs) play important roles in embryogenesis and normal cell growth. The BMP receptors belong to the family of serine/threonine kinase receptors, whose activation has been investigated intensively for the transforming growth factor-beta (TGF-beta) receptor subfamily. However, the interactions between the BMP receptors, the composition of the active receptor complex, and the role of the ligand in its formation have not yet been investigated and were usually assumed to follow the same pattern as the TGF-beta receptors. Here we demonstrate that the oligomerization pattern of the BMP receptors is different and is more flexible and susceptible to modulation by ligand. Using several complementary approaches, we investigated the formation of homomeric and heteromeric complexes between the two known BMP type I receptors (BR-Ia and BR-Ib) and the BMP type II receptor (BR-II). Coimmunoprecipitation studies detected the formation of heteromeric and homomeric complexes among all the BMP receptor types even in the absence of ligand. These complexes were also detected at the cell surface after BMP-2 binding and cross-linking. Using antibody-mediated immunofluorescence copatching of epitope-tagged receptors, we provide evidence in live cells for preexisting heteromeric (BR-II/BR-Ia and BR-II/BR-Ib) and homomeric (BR-II/BR-II, BR-Ia/ BR-Ia, BR-Ib/ BR-Ib, and also BR-Ia/ BR-Ib) oligomers in the absence of ligand. BMP-2 binding significantly increased hetero- and homo-oligomerization (except for the BR-II homo-oligomer, which binds ligand poorly in the absence of BR-I). In contrast to previous observations on TGF-beta receptors, which were found to be fully homodimeric in the absence of ligand, the BMP receptors show a much more flexible oligomerization pattern. This novel feature in the oligomerization mode of the BMP receptors allows higher variety and flexibility in their responses to various ligands as compared with the TGF-beta receptors.

Clustering and signalling of cell receptors

“…This is consistent with a recent experimental finding [18]. We have studied clustering, based on an appropriate definition of clustering correlation function.…”

Osteogenic potential of five different recombinant human bone morphogenetic protein adenoviral vectors in the rat

“…Most proteins of the BMP family bind to two types of signaling receptors termed as type I and type II receptors. 52 The type II receptors are kinases, which are constitutively active without ligand stimulation. Upon ligand binding and formation of type II and type I receptor complexes, followed by possible receptor conformational changes, type I receptors are phosphorylated and activated by type II receptor kinases.…”

Activation of c-Jun NH2-terminal kinase 1 increases cellular responsiveness to BMP-2 and decreases binding of inhibitory Smad6 to the type 1 BMP receptor

“…(8,9) The experimental study of BMP-2 signaling and BMP responsiveness is challenging and is complicated by at least two issues: (1) cross-talk with numerous other pathways (15,16) and (2) lack of synchronization of cellular differentiation in spontaneously differentiating heterogeneous preosteoblast cultures. (25) Several studies have demonstrated that MAPKs are required for BMP-2-induction of the osteoblastic phenotype and mineral deposition in vitro.…”

“…Once activated, R-Smads associate with Smad4 and translocate to the nucleus to regulate gene expression. (8)(9)(10) Inhibitory Smads6 and -7 potently antagonize the BMP/Smad pathway by interacting with activated BMPR-I and preventing activation of R-Smads or by competing with activated R-Smads to form complexes with Smad4. (11)(12)(13)(14) BMP responsiveness may be modulated in part by complex cross-talk with other signaling pathways, including mitogenactivated protein kinases (MAPKs).…”