Identity of urinary trypsin inhibitor-binding protein to link protein

Hiroshi Kobayashi, Yasuyuki Hirashima, Guang Wei Sun, Michio Fujie, Takashi Nishida, Masaharu Takigawa, Toshihiko Terao

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

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 49-kDa protein (UTI-BP40) was purified from the cultured human chondrosarcoma cell line HCS-2/8 by UTI affinity chromatography. Purified UTI- BP40 was digested with trypsin, and the amino acid sequences of the peptide fragments were determined. The sequences of six tryptic fragments of UTI-BP40 were identical to subsequences present in human link protein (LP). Authentic bovine LP and UTI- BP40 displayed identical electrophoretic 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 part 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-BP40 exhibited significant hyaluronic acid binding. These results demonstrate that UTI-BP40 is identical to LP and that the NH2-terminal domain of UTI is involved in the interaction with the NH2-terminal fragment of LP, which is bound to hyaluronic acid in the extracellular matrix.

Original languageEnglish
Pages (from-to)21185-21191
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number28
DOIs
Publication statusPublished - Jul 14 2000

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Carrier Proteins
Hyaluronic Acid
link protein
urinastatin
Affinity chromatography
Chondrosarcoma
Peptide Fragments
Molecules
Protease Inhibitors
Affinity Chromatography
Trypsin
Extracellular Matrix
Amino Acid Sequence
Cells
Amino Acids
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identity of urinary trypsin inhibitor-binding protein to link protein. / Kobayashi, Hiroshi; Hirashima, Yasuyuki; Sun, Guang Wei; Fujie, Michio; Nishida, Takashi; Takigawa, Masaharu; Terao, Toshihiko.

In: Journal of Biological Chemistry, Vol. 275, No. 28, 14.07.2000, p. 21185-21191.

Research output: Contribution to journalArticle

Kobayashi, Hiroshi ; Hirashima, Yasuyuki ; Sun, Guang Wei ; Fujie, Michio ; Nishida, Takashi ; Takigawa, Masaharu ; Terao, Toshihiko. / Identity of urinary trypsin inhibitor-binding protein to link protein. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 28. pp. 21185-21191.
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