Involvement of Disulfide Bond Formation in the Activation of Heparanase

Simizu, Siro; Suzuki, Takehiro; Muroi, Makoto; Lai, Ngit Shin; Takagi, Satoshi; Dohmae, Naoshi; Osada, Hiroyuki

doi:10.1158/0008-5472.can-07-1053

Cited by 46 publications

(32 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To remove SDS, the gel was washed with 2.5% Triton X-100 for 1 h and incubated with incubation buffer [50 mmol/L Tris (pH 7.5), 200 mmol/L NaCl, and 10 mmol/L CaCl 2 ] for 24 h at 37jC. Then, the gel was stained with Coomassie Brilliant Blue G-250 (26). In the case of the cell lysates, cells were washed with PBS and sonicated for 10 s after lysis without 2-mercaptoethanol sampling buffer at 4jC.…”

Section: Methodsmentioning

confidence: 99%

RECK Negatively Regulates Matrix Metalloproteinase-9 Transcription

2009

Self Cite

View full text Add to dashboard Cite

RECK, a glycosylphosphatidylinositol-anchored glycoprotein, inhibits the enzymatic activities of some matrix metalloproteinases (MMP), thereby suppressing tumor cell metastasis; however, the detailed mechanism is still obscure. In this study, we compared the gene expression profiles between mock-and RECK-transfected HT1080 cells and showed that RECK decreases MMP-9 mRNA levels but not other MMP mRNA levels. Moreover, treatment with RECK-specific siRNA increased MMP-9 mRNA in RECK-expressing cells. The promoter assay showed that MMP-9 promoter activity was suppressed by RECK and that RECK-mediated suppression of MMP-9 promoter activity requires 12-O-tetradecanoylphorbol-13-acetate-responsive element (TRE) and KB sites. Moreover, the binding ability of Fra-1 and c-Jun to TRE within the MMP-9 promoter region was suppressed by RECK. Thus, these results show that RECK is a negative regulator of MMP-9 transcription. [Cancer Res 2009;69(4):1502-8]

show abstract

Section: Methodsmentioning

confidence: 99%

RECK Negatively Regulates Matrix Metalloproteinase-9 Transcription

2009

Self Cite

View full text Add to dashboard Cite

show abstract

“…Five N-glycosylation sites, 87 one S-cysteinylation and two disulfide bridges 88 have been characterized (Supporting Information Figure 1S). The N-glycosylation sites are located on the model surface ( Figure 5A), as expected.…”

Section: General Organization Of the Heparanase Structure And Validationmentioning

confidence: 99%

Molecular model of human heparanase with proposed binding mode of a heparan sulfate oligosaccharide and catalytic amino acids

Sapay

Cabannes²,

Petitou³

et al. 2011

Biopolymers

View full text Add to dashboard Cite

Heparan sulfate is abundantly present in the extracellular matrix. As other glycosaminoglycans, it is synthesized in the Golgi apparatus and then exposed on the cell surface. The glucuronidase activity of human heparanase plays a major role in the structural remodeling of the extracellular matrix, which underlies cell migration, hence tumor invasion. Heparanase is therefore a major target for anti-cancer treatment. Several inhibitors of its enzymatic activity have been synthesized. However, their design is limited by the absence of experimental structure of the protein. Homology modeling is proposed based on the structure of the endoxylanase from Penicillium simplicissimum co-crystallized with a series of xylan oligosaccharide. The new heparanase model is consistent with the few experimental data suited for the validation of such work. Furthermore, the presence of natural substrates in the template structure allowed us to propose a binding model for a hydrolyzed heparin sulfate pentasaccharide. Several lysine residues have been identified to play a key role in binding to the anionic polysaccharide substrate. In addition, two phenylalanine residues are also potentially important for the interaction with the substrate. The enzymatic mechanism investigated in the light of this new model allows for the proposal of several amino acids that can influence the protonation state of the nucleophile and the proton donor.

show abstract

“…Control of heparanase post-translational modifications (4,11,12), including proteolytic processing and activation (13)(14)(15)(16), represent important regulatory mechanisms. The human heparanase cDNA has an open reading frame that encodes for 543 amino acids yet the latent proheparanase is 65-kDa in size due to glycosylation (1,11).…”

mentioning

confidence: 99%

Cathepsin L Is Responsible for Processing and Activation of Proheparanase through Multiple Cleavages of a Linker Segment

Abboud-Jarrous

Atzmon

Peretz

et al. 2008

Journal of Biological Chemistry

156

125

View full text Add to dashboard Cite

Heparanase is an endo-␤-D-glucuronidase that degrades heparan sulfate in the extracellular matrix and on the cell surface. Human proheparanase is produced as a latent protein of 543 amino acids whose activation involves excision of an internal linker segment (Ser 110 -Gln 157 ), yielding the active heterodimer composed of 8-and 50-kDa subunits. Applying cathepsin L knock-out tissues and cultured fibroblasts, as well as cathepsin L gene silencing and overexpression strategies, we demonstrate, for the first time, that removal of the linker peptide and conversion of proheparanase into its active 8 ؉ 50-kDa form is brought about predominantly by cathepsin L. Excision of a 10-amino acid peptide located at the C terminus of the linker segment between two functional cathepsin L cleavage sites (Y156Q and Y146Q) was critical for activation of proheparanase. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry demonstrates that the entire linker segment is susceptible to multiple endocleavages by cathepsin L, generating small peptides. Mass spectrometry demonstrated further that an active 8-kDa subunit can be generated by several alternative adjacent endocleavages, yielding the precise 8-kDa subunit and/or slightly elongated forms. Altogether, the mode of action presented here demonstrates that processing and activation of proheparanase can be brought about solely by cathepsin L. The critical involvement of cathepsin L in proheparanase processing and activation offers new strategies for inhibiting the prometastatic, proangiogenic, and proinflammatory activities of heparanase.

show abstract

Involvement of Disulfide Bond Formation in the Activation of Heparanase

Cited by 46 publications

References 50 publications

RECK Negatively Regulates Matrix Metalloproteinase-9 Transcription

RECK Negatively Regulates Matrix Metalloproteinase-9 Transcription

Molecular model of human heparanase with proposed binding mode of a heparan sulfate oligosaccharide and catalytic amino acids

Cathepsin L Is Responsible for Processing and Activation of Proheparanase through Multiple Cleavages of a Linker Segment

Contact Info

Product

Resources

About