Interactions of high affinity insulin-like growth factor-binding proteins with the type V transforming growth factor-P receptor in mink lung epithelial cells
Sm. Leal et al., Interactions of high affinity insulin-like growth factor-binding proteins with the type V transforming growth factor-P receptor in mink lung epithelial cells, J BIOL CHEM, 274(10), 1999, pp. 6711-6717
High affinity insulin-like growth factor-binding proteins (IGFBP-1 to -6) a
re a family of structurally homologous proteins that induce cellular respon
ses by insulin-libe growth factor (IGF)-dependent and -independent mechanis
ms. The IGFBP-3 receptor, which mediates the IGF-independent growth inhibit
ory response, has recently been identified as the type V transforming growt
h factor-beta receptor (T beta R-V) (Leal, S.M, Liu, Q,L,, Huang, S.S., and
Huang, J, S, (1997) J, Biob Chem, 272, 20572-20576), To characterize the i
nteractions of high affinity IGFBPs with T beta R-V, mink lung epithelial c
ells (Mv1Lu cells) were incubated with I-125-labeled recombinant human IGFB
Ps (I-125-IGFBP-1 to -6) in the presence of the cross-linking agent disucci
nimidyl suberate and analyzed by 5% SDS-polyacrylamide gel electrophoresis
and autoradiography. I-125-IGFBP-3, -4, and -5 but not I-125-IGFBP-1, -2, a
nd -6 bound to T beta R-V as demonstrated by the detection of the similar t
o 400-kDa I-125-IGFBP T beta R-V cross-linked complex in the cell lysates a
nd immunoprecipitates. The analyses of 125I-labeled Ligand binding competit
ion and DNA synthesis inhibition revealed that IGFBP-3 was a more potent Li
gand for TPR-V than IGFBP-4 or -5, Most of the high affinity I-125-IGFBPs f
ormed dimers at the cell surface. The cell-surface dimer of I-125-IGFBP-3 p
referentially bound to and was cross-linked to TPR-V in the presence of dis
uccinimidyl suberate, IGFBP-3 did not stimulate the cellular phosphorylatio
n of Smad2 and Smad3, key transducers of the transforming growth factor-bet
a type I/type II receptor (T beta R-IT beta R-II) heterocomplex-mediated si
gnaling. These results suggest that IGFBP-3, -4, and -5 are specific ligand
s for T beta R-V, which mediates the growth inhibitory response through a s
ignaling pathway(s) distinct from that mediated by the T beta R-I and T bet
a R-II heterocomplex.