{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/39271"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/39271","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Isolation and characterization of cathepsin Z, a lysosomal cysteine proteinase","abstract":"Cathepsin Z is a cysteine proteinase found in lysosomes of human cells. It was detected in human cultured cell lines using the peptidyl diazomethane inhibitor Fmoc-Leu-Leu- [¹²⁵]Tyr-CHN₂. The labeling of cathepsin Z by the inhibitor was both time- and concentration-dependent, and the proteinase was found in all human cell lines examined. The characteristics of cathepsin Z were examined in U-937 cells, a human monocytic line. The labeling of cathepsin Z was blocked by pre-incubation of the cells either in non-iodinated inhibitor or in the epoxysuccinyl peptide inhibitor E-64d, a specific inhibitor of cysteine proteinases. Cathepsin Z was not immunoprecipitated by antisera specific for cathepsins B, L, or S. Cathepsin Z has been estimated to be at millimolar concentrations in lysosomes, suggesting that it is a major lysosomal proteinase. The molecular weight of cathepsin Z was calculated to be 22.4 kDa by SDS-PAGE and 45— 47 kDa by native PAGE and gel exclusion chromatography, indicating that it is dimeric. Cathepsin Z is susceptible to digestion by endoglycosidase H, and oligosaccharides comprise 3.1 kDa of the reduced molecular weight. The expression of cathepsin Z was not affected by differentiation of U-937 cells with phorbol ester, unlike the expression of cathepsins B and S. Undifferentiated U-937 cells express low levels of cathepsins B and S; after differentiation, expression of cathepsins B and S is greatly increased. Cathepsin Z was purified from U-937 cells by anion exchange chromatography (Mono Q), affinity chromatography (concanavalin A), and preparatory electrophoresis. N-terminal sequence analysis of both the purified protein and fragments of the protein from a V8 digest indicates that cathepsin Z is a member of the papain superfamily of cysteine proteinases.","abstract_html":"Cathepsin Z is a cysteine proteinase found in lysosomes of human cells. It was detected in human cultured cell lines using the peptidyl diazomethane inhibitor Fmoc-Leu-Leu- [¹²⁵]Tyr-CHN₂. The labeling of cathepsin Z by the inhibitor was both time- and concentration-dependent, and the proteinase was found in all human cell lines examined. The characteristics of cathepsin Z were examined in U-937 cells, a human monocytic line. The labeling of cathepsin Z was blocked by pre-incubation of the cells either in non-iodinated inhibitor or in the epoxysuccinyl peptide inhibitor E-64d, a specific inhibitor of cysteine proteinases. Cathepsin Z was not immunoprecipitated by antisera specific for cathepsins B, L, or S. Cathepsin Z has been estimated to be at millimolar concentrations in lysosomes, suggesting that it is a major lysosomal proteinase. The molecular weight of cathepsin Z was calculated to be 22.4 kDa by SDS-PAGE and 45— 47 kDa by native PAGE and gel exclusion chromatography, indicating that it is dimeric. Cathepsin Z is susceptible to digestion by endoglycosidase H, and oligosaccharides comprise 3.1 kDa of the reduced molecular weight. The expression of cathepsin Z was not affected by differentiation of U-937 cells with phorbol ester, unlike the expression of cathepsins B and S. Undifferentiated U-937 cells express low levels of cathepsins B and S; after differentiation, expression of cathepsins B and S is greatly increased. Cathepsin Z was purified from U-937 cells by anion exchange chromatography (Mono Q), affinity chromatography (concanavalin A), and preparatory electrophoresis. N-terminal sequence analysis of both the purified protein and fragments of the protein from a V8 digest indicates that cathepsin Z is a member of the papain superfamily of cysteine proteinases.","abstract_has_math":false,"creators":["DeCourcy, Kristi R."],"institution":"Virginia Tech","degree_name":"Ph. D.","degree_level":"doctoral","degree_discipline":"Biochemistry and Anaerobic Microbiology","degree_department":"Biochemistry and Anaerobic Microbiology","school":null,"contributors":[],"advisors":[],"committee_chairs":["Mason, Robert W.","Storrie, Brian"],"committee_members":["Bender, Patrick K.","Newton, William E.","Rutherford, Charles L."],"year":1995,"date_issued":"1995-05-05","date_published":"1995-05-05","updated_at":"2026-07-22T22:19:19Z","subjects":["human cell lines"],"languages":["en"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["etd-08272007-163757"],"render_values":[{"text":"etd-08272007-163757","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10919/39271","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Mason, Robert W.","Storrie, Brian"]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Bender, Patrick K.","Newton, William E.","Rutherford, Charles L."]},{"key":"dc:contributor.department","label":"Department","values":["Biochemistry and Anaerobic Microbiology"]},{"key":"dc:creator","label":"Author","values":["DeCourcy, Kristi R."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2014-03-14T21:18:22Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2014-03-14T21:18:22Z","2007-08-27"]},{"key":"dc:date.issued","label":"Date","values":["1995-05-05"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation"]},{"key":"dc:type.dcmitype","label":"Dc Type Dcmitype","values":["Text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry and Anaerobic Microbiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph. 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It was detected in human cultured cell lines using the peptidyl diazomethane inhibitor Fmoc-Leu-Leu- [¹²⁵]Tyr-CHN₂. The labeling of cathepsin Z by the inhibitor was both time- and concentration-dependent, and the proteinase was found in all human cell lines examined. The characteristics of cathepsin Z were examined in U-937 cells, a human monocytic line. The labeling of cathepsin Z was blocked by pre-incubation of the cells either in non-iodinated inhibitor or in the epoxysuccinyl peptide inhibitor E-64d, a specific inhibitor of cysteine proteinases. Cathepsin Z was not immunoprecipitated by antisera specific for cathepsins B, L, or S. Cathepsin Z has been estimated to be at millimolar concentrations in lysosomes, suggesting that it is a major lysosomal proteinase. The molecular weight of cathepsin Z was calculated to be 22.4 kDa by SDS-PAGE and 45— 47 kDa by native PAGE and gel exclusion chromatography, indicating that it is dimeric. Cathepsin Z is susceptible to digestion by endoglycosidase H, and oligosaccharides comprise 3.1 kDa of the reduced molecular weight. The expression of cathepsin Z was not affected by differentiation of U-937 cells with phorbol ester, unlike the expression of cathepsins B and S. Undifferentiated U-937 cells express low levels of cathepsins B and S; after differentiation, expression of cathepsins B and S is greatly increased. Cathepsin Z was purified from U-937 cells by anion exchange chromatography (Mono Q), affinity chromatography (concanavalin A), and preparatory electrophoresis. N-terminal sequence analysis of both the purified protein and fragments of the protein from a V8 digest indicates that cathepsin Z is a member of the papain superfamily of cysteine proteinases."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph. D."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["BTD"]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Isolation and characterization of cathepsin Z, a lysosomal cysteine proteinase"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Mason, Robert W.","Storrie, Brian"],"dc:contributor.committeemember":["Bender, Patrick K.","Newton, William E.","Rutherford, Charles L."],"dc:contributor.department":["Biochemistry and Anaerobic Microbiology"],"dc:creator":["DeCourcy, Kristi R."],"dc:date.accessioned":["2014-03-14T21:18:22Z"],"dc:date.available":["2014-03-14T21:18:22Z","2007-08-27"],"dc:date.issued":["1995-05-05"],"dc:description.abstract":["Cathepsin Z is a cysteine proteinase found in lysosomes of human cells. It was detected in human cultured cell lines using the peptidyl diazomethane inhibitor Fmoc-Leu-Leu- [¹²⁵]Tyr-CHN₂. The labeling of cathepsin Z by the inhibitor was both time- and concentration-dependent, and the proteinase was found in all human cell lines examined. The characteristics of cathepsin Z were examined in U-937 cells, a human monocytic line. The labeling of cathepsin Z was blocked by pre-incubation of the cells either in non-iodinated inhibitor or in the epoxysuccinyl peptide inhibitor E-64d, a specific inhibitor of cysteine proteinases. Cathepsin Z was not immunoprecipitated by antisera specific for cathepsins B, L, or S. Cathepsin Z has been estimated to be at millimolar concentrations in lysosomes, suggesting that it is a major lysosomal proteinase. The molecular weight of cathepsin Z was calculated to be 22.4 kDa by SDS-PAGE and 45— 47 kDa by native PAGE and gel exclusion chromatography, indicating that it is dimeric. Cathepsin Z is susceptible to digestion by endoglycosidase H, and oligosaccharides comprise 3.1 kDa of the reduced molecular weight. The expression of cathepsin Z was not affected by differentiation of U-937 cells with phorbol ester, unlike the expression of cathepsins B and S. Undifferentiated U-937 cells express low levels of cathepsins B and S; after differentiation, expression of cathepsins B and S is greatly increased. Cathepsin Z was purified from U-937 cells by anion exchange chromatography (Mono Q), affinity chromatography (concanavalin A), and preparatory electrophoresis. N-terminal sequence analysis of both the purified protein and fragments of the protein from a V8 digest indicates that cathepsin Z is a member of the papain superfamily of cysteine proteinases."],"dc:description.degree":["Ph. D."],"dc:format.medium":["BTD"],"dc:format.mimetype":["application/pdf"],"dc:identifier.other":["etd-08272007-163757"],"dc:identifier.uri":["http://hdl.handle.net/10919/39271"],"dc:language.iso":["en"],"dc:publisher":["Virginia Tech"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:subject":["human cell lines"],"dc:title":["Isolation and characterization of cathepsin Z, a lysosomal cysteine proteinase"],"dc:type":["Dissertation"],"dc:type.dcmitype":["Text"],"thesis:degree_discipline":["Biochemistry and Anaerobic Microbiology"],"thesis:degree_level":["doctoral"],"thesis:degree_name":["Ph. D."],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:19:19Z"}