Processing of the Human Heparanase Precursor and Evidence That the Active Enzyme Is a Heterodimer

Fairbanks, Michael B.; Mildner, Ana M.; Leone, Joseph W.; Cavey, Gregory S.; Mathews, W R; Drong, Roger F.; Slightom, Jerry L.; Bienkowski, Michael J.; Smith, Clark W.; Bannow, Carol A.; Heinrikson, Robert L.

doi:10.1074/jbc.274.42.29587

Cited by 184 publications

(165 citation statements)

References 24 publications

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“…Regulation at the post-translational level, namely heparanase processing, cellular localization and secretion, has also been implicated as major regulatory mechanisms. [26][27][28]42 Heparanase upregulation has been noted in several primary human tumors, including colon carcinoma by employing in situ hybridization and immunostaining. 31 Moreover, heparanase upregulation in colon carcinoma correlated with increased lymph vessel and vascular infiltration and, most importantly, with reduced postoperative survival rate, 43 suggesting that heparanase expression levels may be considered as an important diagnostic parameter.…”

Section: Discussionmentioning

confidence: 99%

“…30,31 Antibody 1453 was raised in rabbits against the full-length 65 kDa latent heparanase protein. 26 Immunohistochemistry was performed essentially as described. 31,32 Briefly, 5 mm sections were deparaffinized and rehydrated, followed by antigen retrieval (boiling in 10 mM citrate buffer, pH 6, for 10 min in a microwave oven).…”

Section: Spatial and Temporal Heparanase Expressionmentioning

confidence: 99%

“…Fractions (0.2 ml) were eluted with PBS and their radioactivity counted in a l-scintillation counter. Degradation fragments of heparan sulfate side chains generated by heparanase are eluted at 0.5oK av o0.8 (peak II, fractions [15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30]. Nearly intact heparan sulfate proteoglycans are eluted just after the Vo (K av o0.2, peak I, fractions 3-15).…”

Section: Heparanase Activity Assaymentioning

confidence: 99%

“…[26][27][28] Recently, we have generated a polyclonal antibody (733) that preferentially recognizes the 50 kDa heparanase subunit vs the 65 kDa latent enzyme, and used this antibody to study the trafficking routes, processing and localization of heparanase in vitro. 29 Furthermore, this antibody is suitable for staining of paraffin sections.…”

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confidence: 99%

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Spatial and temporal heparanase expression in colon mucosa throughout the adenoma-carcinoma sequence

Doviner

Maly

Kaplan³

et al. 2006

Modern Pathology

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Heparanase is a mammalian endo-b-D-glucuronidase that cleaves heparan sulfate side chains at a limited number of sites. Such enzymatic activity is thought to participate in degradation and remodeling of the extracellular matrix and to facilitate cell invasion associated with tumor metastasis, angiogenesis and inflammation. Traditionally, heparanase activity was well correlated with the metastatic potential of a large number of tumor-derived cell types. More recently, heparanase upregulation has been documented in an increasing number of primary human tumors, correlating with poor postoperative survival and increased tumor vascularity. Here, we employed antiheparanase 733 polyclonal antibody that preferentially recognizes the 50 kDa active heparanase subunit over the 65 kDa proenzyme, as well as anti-heparanase 92.4 monoclonal antibody that recognizes both the latent and the active enzyme, to follow heparanase expression, processing and localization throughout the adenoma-carcinoma transition of the colon epithelium. Normal (nondysplastic) mucosa of the large bowel near epithelial neoplasms, as well as areas of mild dysplasia in adenomas, exhibited a strong reactivity with antibody 733 that became even stronger in foci of moderate dysplasia. Interestingly, although reactivity with antibody 733 was markedly reduced in severe dysplasia and in colorectal carcinoma, response to antibody 92.4 exhibited the opposite trend and staining intensities increased in parallel with tumor stage, the highest being in carcinoma cells. Involvement of latent heparanase (detected by 92.4, but not by 733 antibody) in tumor progression was suggested by activation of the Akt/PKB signal transduction pathway upon heparanase overexpression or exogenous addition to HT29 human colon carcinoma cells. These results suggest that heparanase expression is induced during colon carcinogenesis, and that its processing, conformation and localization are tightly regulated during the course of colon adenoma-carcinoma progression. Heparanase is an endoglycosidase that specifically cleaves heparan sulfate side chains of heparan sulfate proteoglycans. 1,2 Heparan sulfate proteoglycans consist of a protein core to which several heparan sulfate side chains are covalently attached. These complex macromolecules are highly abundant in the extracellular matrix and are thought to play an important structural role, contributing to extracellular matrix integrity and insolubility. 3 In addition, heparan sulfate side chains can bind to a variety of biological mediators such as growth factors, cytokines and chemokines, 4,5 thus functioning as a readily available reservoir that can be liberated upon local or systemic cues. Moreover, heparan sulfate proteoglycans on the cell surface participates directly in signal-transduction cascades by potentiating the interaction between certain growth factors and their receptors. 6-9 Heparan sulfate-degrading activity is thus expected to affect several fundamental aspects of cell behavior under normal and clinical settings. 1,2 Tr...

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Section: Discussionmentioning

confidence: 99%

Section: Spatial and Temporal Heparanase Expressionmentioning

confidence: 99%

Section: Heparanase Activity Assaymentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Spatial and temporal heparanase expression in colon mucosa throughout the adenoma-carcinoma sequence

Doviner

Maly

Kaplan³

et al. 2006

Modern Pathology

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“…It was predicted that the 543-aa polypeptide formed a proenzyme that was processed to be a mature 50 kDa active enzyme after removal of 157 N-terminal amino acids. The active enzyme has been claimed to be a heterodimer that comprises the 50-kDa polypeptide and a short fragment of 8-kDa peptide derived from the N terminus of heparanase proenzyme (11). The active enzyme shows at least 100-fold HS degradation activity in comparison to the proenzyme (8), therefore this processing could be one of the critical regulatory steps of heparanase.…”

mentioning

confidence: 99%

Cell Surface Localization of Heparanase on Macrophages Regulates Degradation of Extracellular Matrix Heparan Sulfate

Sasaki

Higashi

Taka

et al. 2004

The Journal of Immunology

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Extravasation of peripheral blood monocytes through vascular basement membranes requires degradation of extracellular matrix components including heparan sulfate proteoglycans (HSPGs). Heparanase, the heparan sulfate-specific endo-β-glucuronidase, has previously been shown to be a key enzyme in melanoma invasion, yet its involvement in monocyte extravasation has not been elucidated. We examined a potential regulatory mechanism of heparanase in HSPG degradation and transmigration through basement membranes in leukocyte trafficking using human promonocytic leukemia U937 and THP-1 cells. PMA-treated cells were shown to degrade 35S-sulfated HSPG in endothelial extracellular matrix into fragments of an approximate molecular mass of 5 kDa. This was not found with untreated cells. The gene expression levels of heparanase or the enzyme activity of the amount of cell lysates were no different between untreated and treated cells. Immunocytochemical staining with anti-heparanase mAb revealed pericellular distribution of heparanase in PMA-treated cells but not in untreated cells. Cell surface heparanase capped into a restricted area on PMA-treated cells when they were allowed to adhere. Addition of a chemoattractant fMLP induced polarization of the PMA-treated cells and heparanase redistribution at the leading edge of migration. Therefore a major regulatory process of heparanase activity in the cells seems to be surface expression and capping of the enzyme. Addition of the anti-heparanase Ab significantly inhibited enzymatic activity and transmigration of the PMA-treated cells, suggesting that the cell surface redistribution of heparanase is involved in monocyte extravasation through basement membranes.

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Heparanase

Freeman¹,

Hulett²,

Parish³

2002

Wiley Encyclopedia of Molecular Medicine

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Processing of the Human Heparanase Precursor and Evidence That the Active Enzyme Is a Heterodimer

Cited by 184 publications

References 24 publications

Spatial and temporal heparanase expression in colon mucosa throughout the adenoma-carcinoma sequence

Spatial and temporal heparanase expression in colon mucosa throughout the adenoma-carcinoma sequence

Cell Surface Localization of Heparanase on Macrophages Regulates Degradation of Extracellular Matrix Heparan Sulfate

Heparanase

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