Human Inter-α-Trypsin Inhibitor: Localization of the Kunitz-Type Domains in the N-terminal Part of the Molecule and their Release by a Trypsin-Like Proteinase

Reisinger, P. W. M.; Hochstrasser, K; Albrecht, Gerd J.; Lempart, Kathrin; Salier, Jean‐Philippe

doi:10.1515/bchm3.1985.366.1.479

Cited by 43 publications

(9 citation statements)

References 3 publications

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“…Bikunin contains two tandem repeats of Kunitztype domains, and the trypsin inhibitor activity of ITI is localized in this part (23,42). Bikunin is also identified in urine as UTI (urinary trypsin inhibitor), which was shown to be a urine proteoglycan with molecular mass ranging from 40 to 45 kDa (43)(44)(45).…”

Section: Peaksmentioning

confidence: 99%

Evidence for the Covalent Binding of SHAP, Heavy Chains of Inter-α-Trypsin Inhibitor, to Hyaluronan

Zhao

Yoneda

Ohashi

et al. 1995

Journal of Biological Chemistry

166

177

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We previously showed that serum-derived 85-kDa proteins (SHAPs, serum-derived hyaluronan associated proteins) are firmly bound to hyaluronan (HA) synthesized by cultured fibroblasts. SHAPs were then identified to be the heavy chains of inter-␣-trypsin inhibitor (ITI) (Huang, L., Yoneda, M., and Kimata, K. (1993) J. Biol. Chem. 268, 26725-26730). In this study, the SHAP⅐HA complex was isolated from pathological synovial fluid from human arthritis patients. The SHAP⅐HA complex was digested with thermolysin, followed by CsCl gradient centrifugation. The HA-containing fragments thus obtained were further digested with chondroitinase AC II and subjected to TSK gel high performance liquid chromatography (HPLC). Peptide-HA disaccharide-containing fractions (the SHAP⅐HA binding regions) were further purified by reverse phase HPLC. Major peaks were analyzed by protein sequencing and mass spectrometry (electrospray ionization mass spectrometry and collision induced dissociation-MS/MS). By comparison with the reported C-terminal sequences of the human ITI family, the peptides were found to correspond to tetrapeptides derived from the C termini of heavy chains 1 of and 2 of inter-␣-trypsin inhibitor (HC1 and HC2), and heavy chain 3 of pre-␣-trypsin inhibitor (HC3), respectively, and a heptapeptide from HC1. Mass spectrometric analyses suggested that the Cterminal Asp of each heavy chain was esterified to the C6-hydroxyl group of an internal N-acetylglucosamine of HA chain. This report is the first demonstration to give evidence for the covalent binding of proteins to HA.Hyaluronan (HA), 1 has been found as a ubiquitous component of the extracellular matrices of many tissues and in body fluids, including the vitreous body, synovial fluid, lymph, and blood (1-4). It has been suggested that HA plays an important role in many biological processes, such as gamete maturation, tissue morphogenesis, cell migration, and cell proliferation (5-8). HA is also involved in angiogenesis, wound healing, tumor invasion, and pathophysiological responses of tissues to inflammation (9 -12).With regard to functional importance, a large number of HA-binding proteins have been reported, an important subset of which have highly homologous sequences for HA binding. These include link proteins (13), hyaluronectin (14), glial HAbinding protein (15), HA-binding proteoglycan such as aggrecan, PG-M/versican (16), and CD44 (17). These are the proteoglycan tandem repeat families of HA-binding proteins. CD44 is a typical example of the family. Variant forms of CD44 generated by alternative splicing may have individual functions such as lymphocyte homing and tumor cell metastasis (18,19). Tumor necrosis factor-stimulated gene-6, another new member of this family, is tumor necrosis factor or interleukin-1-inducible and was recently shown to bind covalently to inter-␣-trypsin inhibitor (ITI) (20).We previously showed that serum-derived HA-associated proteins (SHAPs) appear to bind covalently to HA (21), and therefore to mediate the binding of HA to cell surfa...

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Section: Peaksmentioning

confidence: 99%

Evidence for the Covalent Binding of SHAP, Heavy Chains of Inter-α-Trypsin Inhibitor, to Hyaluronan

Zhao

Yoneda

Ohashi

et al. 1995

Journal of Biological Chemistry

166

177

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“…As shown in figure 3, 125I-UTI in the plasma was excreted into the urine in its intact form. Plasma ASTI is assumed to be a precursor form of UTI based on its immuno logical and physicochemical properties and NH2-terminal amino acid sequence [14][15][16]. The activity of plasma ASTI in renal failure patients was found to be about 20-fold higher than that of normal subjects [14].…”

Section: Excretion Of !25i-labeled Utimentioning

confidence: 99%

Clearance and Distribution of Acid-Stable Trypsin Inhibitor (ASTI)

Sugiki

Sumi

Maruyama

et al. 1989

Enzyme

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The clearance, organ distribution and metabolic pathway of the acid-stable trypsin inhibitor (ASTI) were studied in mice using 125I-labeled urinary trypsin inhibitor (UTI), the most typical ASTI in the urine. Following intravenous injection of 125I-UTI, the radioactivity disappeared rapidly from the circulation with a half-life of 4 min for the initial part of the curve. Gel filtration of plasma samples revealed that the rapid disappearance of the radioactivity was due to elimination of free inhibitor from the plasma. 125I-UTI was cleared primarily in the kidney. Gel filtration of urine samples showed that part of the radioactivity in the urine appeared at the same elution volume as 125I-UTI in the plasma, indicating that the origin of UTI was ASTI in the plasma.

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“…Further characterization of cDNAs has shown that ITI is composed of distinct heavy chains (heavy chains 1 and 2) that are linked limited proteolysis of ITI in the N-terminal region of the polypeptide chain [4]. UTI is increased in several diseases, including inflammation and cancer [5].…”

Section: Introductionmentioning

confidence: 99%

“…More indications regarding the relationship between ITI, UTI, and other polypeptides derived from them have been given elsewhere [3]. The structure of these molecules, their polypeptidic chains, their proteoglycan moiety, the location of proteinase inhibitory sites, as well as the molecular weights are described elsewhere [1][2][3][4]7].…”

Section: Introductionmentioning

confidence: 99%

Urinary trypsin inhibitor (UTI) and fragments derived from UTI by limited proteolysis efficiently inhibit tumor cell invasion

et al. 1994

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We investigated the effects of purified human urinary trypsin inhibitor (UTI) and fragments derived from UTI by proteolysis on the invasive potential of ovarian cancer cells (HOC-I) and gestational choriocarcinoma cells (SMT-ccl) using an in vitro reconstituted basement membrane invasion assay. These cells express cell-associated plasmin and functional uPA receptors that are partially occupied by ligands. SMT-ccl cells, which express threefold higher levels of cell-associated plasmin activity than HOC-I cells, showed approximately twofold increase in their invasive potential. For the invasion assay, HOC-I cells were primed with exogenous plasminogen, but SMT-ccl cells were not. Human leukocyte elastase (HLE)-digested UTI (22 kDa fragment; UTI-22) inhibited plasmin practically with the same strength as native UTI. Trypsin-digested UTI (20 kDa fragment; UTI-20), however, did not inhibit plasmin significantly. Treatment of cells with UTI or UTI-22 reduced the incidence of tumor cell invasive capacity, whereas the inhibitory effect of UTI-20 was not remarkable. The inhibitory effect on tumor cell invasion was dose-dependent and non-toxic; moreover, it was not mediated by inhibition of the tumor cell chemotactic response or of cell attachment to matrigel. These results indicate that inhibition of the proteolytic enzyme plasmin specifically reduced the invasive capacity of tumor cells in vitro.

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Human Inter-α-Trypsin Inhibitor: Localization of the Kunitz-Type Domains in the N-terminal Part of the Molecule and their Release by a Trypsin-Like Proteinase

Cited by 43 publications

References 3 publications

Evidence for the Covalent Binding of SHAP, Heavy Chains of Inter-α-Trypsin Inhibitor, to Hyaluronan

Evidence for the Covalent Binding of SHAP, Heavy Chains of Inter-α-Trypsin Inhibitor, to Hyaluronan

Clearance and Distribution of Acid-Stable Trypsin Inhibitor (ASTI)

Urinary trypsin inhibitor (UTI) and fragments derived from UTI by limited proteolysis efficiently inhibit tumor cell invasion

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