{"entity": "journal", "iuid": "0a1c01ceff3546a593fc0072ea2ece79", "timestamp": "2026-08-26T22:47:02.205Z", "links": {"self": {"href": "https://publications-affiliated.scilifelab.se/journal/BMC%20Mol%20Cell%20Biol.json"}, "display": {"href": "https://publications-affiliated.scilifelab.se/journal/BMC%20Mol%20Cell%20Biol"}}, "title": "BMC Mol Cell Biol", "issn": "2661-8850", "issn-l": null, "publications_count": 2, "publications": [{"entity": "publication", "iuid": "d5e23d4fea2f420985ef91d8d38ef9ea", "links": {"self": {"href": "https://publications-affiliated.scilifelab.se/publication/d5e23d4fea2f420985ef91d8d38ef9ea.json"}, "display": {"href": "https://publications-affiliated.scilifelab.se/publication/d5e23d4fea2f420985ef91d8d38ef9ea"}}, "title": "Probing contacts of inhibitor locked in transition states in the catalytic triad of DENV2 type serine protease and its mutants by 1H, 19F and 15 N NMR spectroscopy.", "authors": [{"family": "Agback", "given": "Peter", "initials": "P", "orcid": "0000-0003-2226-0746", "researcher": {"href": "https://publications-affiliated.scilifelab.se/researcher/eae6fd83468f4ddfbbb7609d710dec6c.json"}}, {"family": "Woestenenk", "given": "Esmeralda", "initials": "E"}, {"family": "Agback", "given": "Tatiana", "initials": "T"}], "type": "journal article", "published": "2020-05-25", "journal": {"title": "BMC Mol Cell Biol", "issn": "2661-8850", "volume": "21", "issue": "1", "pages": "38", "issn-l": null}, "abstract": "Detailed structural knowledge of enzyme-inhibitor complexes trapped in intermediate state is the key for a fundamental understanding of reaction mechanisms taking place in enzymes and is indispensable as a structure-guided drug design tool. Solution state NMR uniquely allows the study of active sites of enzymes in equilibrium between different tautomeric forms. In this study 1H, 19F and 15 N NMR spectroscopy has been used to probe the interaction contacts of inhibitors locked in transition states of the catalytic triad of a serine protease. It was demonstrated on the serotype II Dengue virus NS2B:NS3pro serine protease and its mutants, H51N and S135A, in complex with high-affinity ligands containing trifluoromethyl ketone (tfk) and boronic groups in the C-terminal of tetra-peptides.\n\nMonitoring 19F resonances, shows that only one of the two isomers of the tfk tetra-peptide binds with NS2B:NS3pro and that access to the bulk of the active site is limited. Moreover, there were no bound water found in proximity of the active site for any of the ligands manifesting in a favorable condition for formation of low barrier hydrogen bonds (LBHB) in the catalytic triad. Based on this data we were able to identify a locked conformation of the protein active site. The data also indicates that the different parts of the binding site most likely act independently of each other.\n\nOur reported findings increases the knowledge of the detailed function of the catalytic triad in serine proteases and could facilitate the development of rational structure based inhibitors that can selectively target the NS3 protease of Dengue type II (DENV2) virus. In addition the results shows the usefulness of probing active sites using 19F NMR spectroscopy.", "doi": "10.1186/s12860-020-00283-0", "pmid": "32450796", "labels": [], "xrefs": [{"db": "pmc", "key": "PMC7249419"}, {"db": "pii", "key": "10.1186/s12860-020-00283-0"}], "notes": [], "created": "2026-08-21T12:37:38.565Z", "modified": "2026-08-21T12:37:38.620Z"}, {"entity": "publication", "iuid": "d44f22a511394066bd8d20dac0c28c93", "links": {"self": {"href": "https://publications-affiliated.scilifelab.se/publication/d44f22a511394066bd8d20dac0c28c93.json"}, "display": {"href": "https://publications-affiliated.scilifelab.se/publication/d44f22a511394066bd8d20dac0c28c93"}}, "title": "Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells.", "authors": [{"family": "Reithmeier", "given": "Anja", "initials": "A"}, {"family": "Norg\u00e5rd", "given": "Maria", "initials": "M"}, {"family": "Ek-Rylander", "given": "Barbro", "initials": "B"}, {"family": "N\u00e4reoja", "given": "Tuomas", "initials": "T", "orcid": "0000-0002-2174-4434", "researcher": {"href": "https://publications-affiliated.scilifelab.se/researcher/2817f45be8464e75a1762c8e379583f1.json"}}, {"family": "Andersson", "given": "G\u00f6ran", "initials": "G"}], "type": "journal article", "published": "2020-03-18", "journal": {"title": "BMC Mol Cell Biol", "issn": "2661-8850", "volume": "21", "issue": "1", "pages": "15", "issn-l": null}, "abstract": "Tartrate-resistant acid phosphatase (TRAP/ ACP5) belongs to the binuclear metallophosphatase family and is present in two isoforms. The primary translation product is an uncleaved TRAP 5a isoform with low phosphatase activity. TRAP 5a can be post-translationally processed to a cleaved TRAP 5b isoform with high phosphatase activity by e.g. cysteine proteinases, such as Cathepsin K (CtsK). The relevance of the phosphatase activity of TRAP 5b has been demonstrated for proliferation, migration and invasion of cancer cells. TRAP-overexpressing MDA-MB-231 breast cancer cells displayed higher levels of TRAP 5a and efficient processing of TRAP 5a to TRAP 5b protein, but no changes in levels of CtsK when compared to mock-transfected cells. In TRAP-overexpressing cells colocalization of TRAP 5a and proCtsK was augmented, providing a plausible mechanism for generation of TRAP 5b. CtsK expression has been associated with cancer progression and has been pharmacologically targeted in several clinical studies.\n\nIn the current study, CtsK inhibition with MK-0822/Odanacatib did not abrogate the formation of TRAP 5b, but reversibly increased the intracellular levels of a N-terminal fragment of TRAP 5b and reduced secretion of TRAP 5a reversibly. However, MK-0822 treatment neither altered intracellular TRAP activity nor TRAP-dependent cell migration, suggesting involvement of additional proteases in proteolytic processing of TRAP 5a. Notwithstanding, CtsK was shown to be colocalized with TRAP and to be involved in the regulation of secretion of TRAP 5a in a breast cancer cell line, while it still was not essential for processing of TRAP 5a to TRAP 5b isoform.\n\nIn cancer cells multiple proteases are involved in cleaving TRAP 5a to high-activity phosphatase TRAP 5b. However, CtsK-inhibiting treatment was able to reduce secretion TRAP 5a from TRAP-overexpressing cancer cells.", "doi": "10.1186/s12860-020-00253-6", "pmid": "32188406", "labels": [], "xrefs": [{"db": "pmc", "key": "PMC7081696"}, {"db": "pii", "key": "10.1186/s12860-020-00253-6"}], "notes": [], "created": "2026-08-21T12:37:36.363Z", "modified": "2026-08-21T12:37:36.489Z"}], "created": "2026-08-21T12:37:36.441Z", "modified": "2026-08-21T12:37:36.441Z"}