Evidence for inhibitors of the cell-surface protease guanidinobenzoatase

Steven, F. S.; Griffin, Margaret M.; Wong, Theresia L. H.; Brown, Bernard S.; Jeffrey, I

doi:10.1042/bst0141290

Cited by 4 publications

(9 citation statements)

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“…Similar results to those shown in Figure 2 were obtained when formaldehyde-fixed tissues in wax embedded sections were stained with 9-aminoacridine. It was previously shown that a similar situation was found in pancreatic acinar cells (Steven et al, 1986c) and it was demonstrated that formaldehyde displaced an inhibitor from the guanidinobenzoatase. Affinity systems were used to isolate this protein Frozen Section (Fig.…”

Section: Resultsmentioning

confidence: 84%

“…Affinity systems were used to isolate this protein Frozen Section (Fig. 1 inhibitor from pancreatic and liver tissue; the purified inhibitor was then exchanged with the cell surface bound inhibitor (Steven et al, 1986c). The evidence presented in this study clearly shows that the colonic tumour cells in vivo are unable to bind 9-aminoacridine due to the presence of an inhibitor which can be displaced by formaldehyde without interfering with the enzyme's ability to bind 9-aminoacridine (Figure 2).…”

Section: Resultsmentioning

confidence: 99%

“…The evidence is presented in the form of fluorescent micrographs which have been obtained after treating the sections with 9-aminoacridine, a successful probe for the active centre of guanidinobenzoatase (Steven et al, 1985(Steven et al, , 1986a. Tumour cells with active enzyme fluoresce yellow (Steven et al, 1985(Steven et al, , 1986c surface receptors or to any specific molecule with an affinity for low molecular weight ligands.…”

mentioning

confidence: 99%

“…This paper concerns a protease which is associated with cells capable of migration and in particular tumour cells (Steven et al, 1985(Steven et al, , 1986a(Steven et al, , b, 1987Johnson et al, 1986). The enzyme is referred to as guanidinobenzoatase (Steven & AlAhmad, 1983), has been purified by affinity chromatography (Steven et al, 1986b) and has been shown to be a protease, cleaving the arginyl bond in the peptide GlyArgGlyAsp (Steven et al, 1986c) considered to be the link region for attachment of cells to fibronectin (Piersbacher & Ruoslahti, 1984). Previous studies have been concerned with the kinetics of inhibition of guanidinobenzoatase (Steven et al, 1985;Steven & Al-Ahmad, 1983) in solution and the application of these data to the selection of fluorescent competitive inhibitors which may be used to locate cells possessing this enzyme (Steven et al, 1985(Steven et al, , 1986a in formaldehyde fixed, wax embedded sections and resin embedded sections (Steven et al, 1986b).…”

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confidence: 99%

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The inhibition of a tumour cell surface protease in vivo and its re-activation by oxidation

Steven

Ali²,

Griffin³

1988

Br J Cancer

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Summary Colonic tumour cells possess a cell surface protease capable of binding 9-aminoacridine to its active centre, thus locating cells when viewed under a fluorescence microscope. In vivo and in frozen sections, the enzyme is masked by a protein inhibitor. This inhibitor can be displaced by formaldehyde fixation of the tissue and then replaced by adding a fresh extract of colon or lung tissue. The inhibitor is modified by oxidation; provided by air, oxidized glutathione or potassium permanganate, resulting in a change in conformation in the inhibitor and this then results in the enzyme binding the fluorescent probe. The effect of oxidation can be reversed by dithiothreitol. It is proposed that these changes are brought about by a disulphide exchange acting on the inhibitor which indirectly controls the activity of the cell surface enzyme in vivo.The steps described above can be conveniently followed on sections of tissue mounted on a microscope slide; this has the advantage that the same cells can be monitored during a sequence of reactions. It is believed that these techniques could well be applied to other enzyme systems than the tumour protease described in this study.

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Section: Resultsmentioning

confidence: 84%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The inhibition of a tumour cell surface protease in vivo and its re-activation by oxidation

Steven

Ali²,

Griffin³

1988

Br J Cancer

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“…This protease can degrade fibronectin (Steven et al, 1986b) and is associated with cells capable of migration (Steven et al, 1985). The cell enzyme exists in multiple forms which show tissue specific inhibition (Steven et al, 1988a).…”

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confidence: 99%

A study of guanidinobenzoatase during development of mesothelioma induced in the rat by fibrous erionite

Steven

Hill²

1988

Br J Cancer

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Characterization of the guanidinobenzoatase in the epididymal fluids of the mouse

et al. 1997

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Guanidinobenzoatase (GB), a proteolytic enzyme found in the epididymal fluids of mice, was purified to apparent homogeneity by molecular sieving and affinity chromatography. It has a molecular mass of 71 kDa and its enzymatic activity is heat labile and sensitive to EGTA. Its kinetic parameters (Km of 6.66 μM and a Vmax of 4.38 nmol/min/mg) were determined using 4‐methylumbelliferyl‐p‐guanidinobenzoate (MUGB) as the substrate. GB activity is concentrated in the cauda epididymal region of the genital tract. Heat‐solubilized whole zonae, biologically active ZP3, and several serine proteinase inhibitors, including a proteinase inhibitor endogenous to the male genital tract, effectively block the ability of GB to hydrolyze MUGB. Pretreating cumulus‐free, zonae intact oocytes with purified GB reduces, in a concentration‐dependent manner, the number of sperm able to bind to the zonae. The function of the soluble enzyme is not known. Its ability to bind both trypsin inhibitors and ZP3 suggests a possible role in gamete recognition. Mol. Reprod. Dev. 47:204–209, 1997. © 1997 Wiley‐Liss, Inc.

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Evidence for inhibitors of the cell-surface protease guanidinobenzoatase

Cited by 4 publications

References 0 publications

The inhibition of a tumour cell surface protease in vivo and its re-activation by oxidation

The inhibition of a tumour cell surface protease in vivo and its re-activation by oxidation

A study of guanidinobenzoatase during development of mesothelioma induced in the rat by fibrous erionite

Characterization of the guanidinobenzoatase in the epididymal fluids of the mouse

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