Type Ialpha collagen is an IGFBP-3 binding protein.

Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) possesses both growth-inhibitory and -potentiating effects on cells, which are independent of IGF action and mediated through specific IGFBP-3 binding proteins/receptors located at the cell membrane, cytosol, or nuclear compartments as well as in the extracellular matrix. We here characterized type Ialpha collagen as one of these IGFBP-3 binding proteins. Human serum was fractionated over an IGFBP-3 affinity column, and bands at 70-100 kDa were eluted as IGFBP-3 ligands. The 100-kDa band was extracted, subjected to N-terminal amino acid sequencing, and identified through database searching as the N-terminal chain of type Ialpha collagen protein. In a separate screening approach, using a yeast two-hybrid system, we cloned the type Ialpha collagen cDNA from a human liver cDNA library as an IGFBP-3 protein partner. Anti-IGFBP-3 antibodies co-immunoprecipitated type Ialpha collagen and IGFBP-3 from the conditioned media of human fibroblasts and vice versa. We demonstrated through ligand dot blot analysis that type Ialpha collagen binds IGFBP-3. IGFBP-3 mutants, with altered sequence at the nuclear localization sequence, bound type Ialpha collagen poorly. Western immunoblot showed that type Ialpha collagen binds only IGFBP-3 but not IGF-I, suggesting an IGF-I-independent mechanism of this interaction. Physiological effects of IGFBP-3-collagen interactions may include modulation of cell adhesion and migration.