Identity of urinary trypsin inhibitor-binding protein to link protein

Urinary trypsin inhibitor (UTI), a Kunitz-type protease inhibitor, directly binds to some types of cells via cell-associated UTI-binding proteins (UTI -BPs). Here we report that the 40-kDa protein (UTI-BP40) was purified from the cultured human chondrosarcoma cell line HCS-2/8 by UTI affinity chromat ography. Purified UTI-BP40 was digested with trypsin, and the amino acid se quences of the peptide fragments were determined. The sequences of six tryp tic fragments of UTI-BP40 were identical to subsequences present in human l ink protein (LP). Authentic bovine LP and UTI-BP40 displayed identical elec trophoretic and chromatographic behavior. The UTI-binding properties of UTI -BP40 and LP were indistinguishable. Direct binding and competition studies strongly demonstrated that the NH2-terminal fragment is the UTI-binding pa rt of the LP molecule, that the COOH-terminal UTI fragment (HI-8) failed to bind the NH2-terminal subdomain of the LP molecule, and that LP and UTI-BP 40 exhibited significant hyaluronic acid binding. These results demonstrate that UTI-BP40 is identical to LP and that the NH2-terminal domain of UTI i s involved in the interaction with the NH2-terminal fragment of LP, which i s bound to hyaluronic acid in the extracellular matrix.