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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Insights into cathepsin-B activity in mature dentin matrix

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Carrilho, Marcela R. [1] ; Scaffa, Polliana [2] ; Oliveira, Vitor [3] ; Tjaderhane, Leo [4, 5, 6] ; Tersariol, Ivarne L. [7] ; Pashley, David H. [8] ; Tay, Franklin [8] ; Nascimento, Fabio D. [9]
Total Authors: 8
[1] Midwestern Univ, Coll Dent Med Illinois, 555 31st St, SH 211-X, Downers Grove, IL 60515 - USA
[2] Oregon Hlth & Sci Univ, Biomat & Biomech, Portland, OR 97201 - USA
[3] Univ Fed Sao Paulo, Dept Biophys, Sao Paulo - Brazil
[4] Univ Oulu, Oulu - Finland
[5] Univ Helsinki, Helsinki Univ Hosp, Dept Oral & Maxillofacial Dis, Helsinki - Finland
[6] Oulu Univ Hosp, Med Res Ctr Oulu MRC Oulu, Res Unit Oral Hlth Sci, Oulu - Finland
[7] Univ Fed Sao Paulo, Dept Biochem, Sao Paulo - Brazil
[8] Augusta Univ, Dent Coll Georgia, Augusta, GA - USA
[9] Univ Mogi das Cruzes, Interdisciplinary Ctr Biochem Invest, Mogi Das Cruzes, SP - Brazil
Total Affiliations: 9
Document type: Journal article
Source: ARCHIVES OF ORAL BIOLOGY; v. 117, SEP 2020.
Web of Science Citations: 0

Objective: Cysteine proteases are lysosomal enzymes that, under specific circumstances, may be secreted into the extracellular space and participate in protein turnover. This study investigated the involvement of cathepsin B in the gelatinolytic activity of mature dentin matrices at neutral pH. Design: Human dentin fragments were made into powder and enzymes were extracted using guanidine-HCl/EDTA. Host-derived dentin proteases (cathepsin B, MMP-2 and MMP-9) were identified by immunoblotting, and their activities were evaluated spectrofluorimetrically using fluorogenic substrates. Proteases activities were monitored by measuring the rate of hydrolysis of substrates in the presence/absence of MMP- or cysteine cathepsin inhibitors, at neutral pH (7.4). Mass spectroscopy was used to determine the substrates' cleavage points. Reverse zymography was performed to examine the gelatinolytic activity of cathepsin B. Results: Western-blots of dentin extracts yielded strong bands at 95, 72 and 30 kDa, corresponding respectively to MMP-9, MMP-2 and Cathepsin B. Greater fluorogenic substrates hydrolysis occurred in the absence of MMP and cysteine cathepsin inhibitors than in their presence. Cathepsin B exhibited significant gelatinolytic activity. Conclusions: Together with MMP-2 and MMP-9, cathepsin B also account for the host-derived gelatinolytic activity and matrix turnover of mature dentin at physiological, neutral pH. (AU)

Grantee:Marcela Rocha de Oliveira Carrilho
Support type: Regular Research Grants
FAPESP's process: 13/05822-9 - Proteinase-Activated Receptors (PARs) in the dentin-pulp complex: identification, modulation and signal tranduction in caries disease
Grantee:Fábio Dupart Nascimento
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 11/12226-8 - Activity of host-derived proteases on degradation of human dentin treated for bonding procedures
Grantee:Marcela Rocha de Oliveira Carrilho
Support type: Scholarships abroad - Research