Spontaneous formation of a proteolytic B1 and B2 bradykinin receptor complex with enhanced signaling capacity.
B1 bradykinin receptor (B1R) induction is critical in the adaptation of the kinin-mediated inflammatory response from a B2 bradykinin receptor (B2R) subtype to a B1R subtype that occurs during chronic insult. Here, we show that B1R spontaneously forms a proteolytic plasma membrane complex with B2R along with increased receptor signaling capacity. Co-expression of hemagglutinin-tagged B2R with FLAG-tagged B1R in HEK293 cells resulted in degradation of B2R as determined by the diminution of the intact 65-kDa B2R species and the appearance of proteolytic B2R products at 30-40 kDa and by the reduction in B2R bradykinin binding sites. On the other hand, the 35-kDa B1R remained intact. Receptor co-expression also led to an increase in constitutive and agonist-stimulated receptor signaling. Selective immunoprecipitation with epitope-specific antibodies revealed a spontaneously formed heterologous receptor complex, which was composed of the intact 35-kDa B1R and the B2R degradation products. Cellular fractionation, cell surface biotinylation, and immunoelectron microscopy showed that B2R.B1R complexes were present on the cell surface. This is the first evidence that a heterologous G protein-coupled receptor complex in the plasma membrane is linked to proteolytic degradation of a participating receptor, and this mechanism may contribute to the adaptation of the kinin response from a B2 type to a B1 type during chronic insult.
Kang
Dong Soo
DS
Department of Physiological Sciences, Lund University, Lund SE-22184, Sweden.
Ryberg
Kristina
K
Mörgelin
Matthias
M
Leeb-Lundberg
L M Fredrik
LM
D019921:Biotinylation; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D000939:Epitopes; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D016253:Microscopy, Immunoelectron; D011233:Precipitin Tests; D011485:Protein Binding; D043783:Receptor, Bradykinin B1 / Q000737:chemistry*; D043782:Receptor, Bradykinin B2 / Q000737:chemistry*; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D018076:DNA, Complementary; D000939:Epitopes; D043783:Receptor, Bradykinin B1; D043782:Receptor, Bradykinin B2