Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage
We used embryonic skeletal cartilage known to have high levels of alkaline phosphatase activity to determine whether growing cartilage has phosphotyrosine phosphatase activity and phosphotyrosinyl histone phosphatase activity at physiologc pH. Embryonic chick pelvic cartilage and fetal pig scapular...
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| Veröffentlicht in: | Metabolism, clinical and experimental clinical and experimental, 1985-02, Vol.34 (2), p.169-175 |
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| creator | Burch, Warner M. Hamner, Gail Wuthier, Roy E. |
| description | We used embryonic skeletal cartilage known to have high levels of alkaline phosphatase activity to determine whether growing cartilage has phosphotyrosine phosphatase activity and phosphotyrosinyl histone phosphatase activity at physiologc pH. Embryonic chick pelvic cartilage and fetal pig scapular growth-plate cartilage were assayed using phosphotyrosine as substrate at pH 7.5 and the amount of tyrosine generated measured. Both cartilage models had K
m for phosphotyrosine between 6 to 24 μmol/L. Phosphotyrosine phosphatase activity correlated with alkaline phosphatase activity as assessed by (1) distribution of histologic staining for alkaline phosphatase within the cartilages, (2) hormonal stimulation of cartilage alkaline phosphatase activity in vitro, (3) comparison of alkaline phosphatase and phosphotyrosine phosphatase activities in the presence of known inhibitors (vanadate, levamisole, homoarginine, and zinc), and (4) assaying chick epiphyseal cartilage alkaline phosphatase purified to homogeneity for phosphotyrosine phosphatase activity. Areas of cartilage with elevated alkaline phosphatase activity also had raised phosphotyrosine phosphatase activity. Triiodothyronine, a known stimulator of cartilage alkaline phosphatase, increased chick cartilage alkaline phosphatase activity 88% and phosphotyrosine phosphatase activity 106%, and stimulated porcine growth-plate cartilage alkaline phosphatase activity 91% and phosphotyrosine phosphatase activity 145% after 3 days of in vitro incubation. Each of the inhibitors block alkaline phosphatase and phosphotyrosine phosphatase activities. The purified alkaline phosphatase had a K
m for phosphotyrosine of 18 μmol/L and V
max of 5700 nmol tyrosine/mg protein/h, which is well over 1000-fold higher than the phosphotyrosine phosphatase activity found in the above preparations of pelvic and scapular cartilage. Furthermore, cartilage alkaline phosphatase dephosphorylated phosphotyrosinated histone and had little to no activity when histones phosphorylated at serine and threonine were used as substrate. Our studies support the hypothesis that alkaline phosphatase functions as a neutral phosphoprotein phosphatase, ie, a phosphotyrosine phosphatase, and is involved in the regulation of protein phosphorylation-dephosphorylation reactions. |
| doi_str_mv | 10.1016/0026-0495(85)90128-3 |
| format | Article |
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m for phosphotyrosine between 6 to 24 μmol/L. Phosphotyrosine phosphatase activity correlated with alkaline phosphatase activity as assessed by (1) distribution of histologic staining for alkaline phosphatase within the cartilages, (2) hormonal stimulation of cartilage alkaline phosphatase activity in vitro, (3) comparison of alkaline phosphatase and phosphotyrosine phosphatase activities in the presence of known inhibitors (vanadate, levamisole, homoarginine, and zinc), and (4) assaying chick epiphyseal cartilage alkaline phosphatase purified to homogeneity for phosphotyrosine phosphatase activity. Areas of cartilage with elevated alkaline phosphatase activity also had raised phosphotyrosine phosphatase activity. Triiodothyronine, a known stimulator of cartilage alkaline phosphatase, increased chick cartilage alkaline phosphatase activity 88% and phosphotyrosine phosphatase activity 106%, and stimulated porcine growth-plate cartilage alkaline phosphatase activity 91% and phosphotyrosine phosphatase activity 145% after 3 days of in vitro incubation. Each of the inhibitors block alkaline phosphatase and phosphotyrosine phosphatase activities. The purified alkaline phosphatase had a K
m for phosphotyrosine of 18 μmol/L and V
max of 5700 nmol tyrosine/mg protein/h, which is well over 1000-fold higher than the phosphotyrosine phosphatase activity found in the above preparations of pelvic and scapular cartilage. Furthermore, cartilage alkaline phosphatase dephosphorylated phosphotyrosinated histone and had little to no activity when histones phosphorylated at serine and threonine were used as substrate. Our studies support the hypothesis that alkaline phosphatase functions as a neutral phosphoprotein phosphatase, ie, a phosphotyrosine phosphatase, and is involved in the regulation of protein phosphorylation-dephosphorylation reactions.</description><identifier>ISSN: 0026-0495</identifier><identifier>EISSN: 1532-8600</identifier><identifier>DOI: 10.1016/0026-0495(85)90128-3</identifier><identifier>PMID: 2982079</identifier><language>eng</language><publisher>New York, NY: Elsevier Inc</publisher><subject>Alkaline Phosphatase - antagonists & inhibitors ; Alkaline Phosphatase - metabolism ; Animals ; Biological and medical sciences ; Cartilage - drug effects ; Cartilage - embryology ; Cartilage - enzymology ; Chick Embryo ; Electrophoresis, Polyacrylamide Gel ; Fetus - enzymology ; Fundamental and applied biological sciences. Psychology ; Growth Plate - enzymology ; Histocytochemistry ; Histones - metabolism ; Hydrogen-Ion Concentration ; In Vitro Techniques ; Kinetics ; Phosphoprotein Phosphatases - antagonists & inhibitors ; Phosphoprotein Phosphatases - metabolism ; Phosphoric Monoester Hydrolases - antagonists & inhibitors ; Phosphoric Monoester Hydrolases - metabolism ; Skeleton and joints ; Swine - embryology ; Triiodothyronine - pharmacology ; Vertebrates: osteoarticular system, musculoskeletal system</subject><ispartof>Metabolism, clinical and experimental, 1985-02, Vol.34 (2), p.169-175</ispartof><rights>1985</rights><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c483t-5b4e6a07939ea430609a6d492f3b679a390a7905a33f1dd745909cdc21a4c4e13</citedby><cites>FETCH-LOGICAL-c483t-5b4e6a07939ea430609a6d492f3b679a390a7905a33f1dd745909cdc21a4c4e13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0026049585901283$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9274134$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2982079$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Burch, Warner M.</creatorcontrib><creatorcontrib>Hamner, Gail</creatorcontrib><creatorcontrib>Wuthier, Roy E.</creatorcontrib><title>Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage</title><title>Metabolism, clinical and experimental</title><addtitle>Metabolism</addtitle><description>We used embryonic skeletal cartilage known to have high levels of alkaline phosphatase activity to determine whether growing cartilage has phosphotyrosine phosphatase activity and phosphotyrosinyl histone phosphatase activity at physiologc pH. Embryonic chick pelvic cartilage and fetal pig scapular growth-plate cartilage were assayed using phosphotyrosine as substrate at pH 7.5 and the amount of tyrosine generated measured. Both cartilage models had K
m for phosphotyrosine between 6 to 24 μmol/L. Phosphotyrosine phosphatase activity correlated with alkaline phosphatase activity as assessed by (1) distribution of histologic staining for alkaline phosphatase within the cartilages, (2) hormonal stimulation of cartilage alkaline phosphatase activity in vitro, (3) comparison of alkaline phosphatase and phosphotyrosine phosphatase activities in the presence of known inhibitors (vanadate, levamisole, homoarginine, and zinc), and (4) assaying chick epiphyseal cartilage alkaline phosphatase purified to homogeneity for phosphotyrosine phosphatase activity. Areas of cartilage with elevated alkaline phosphatase activity also had raised phosphotyrosine phosphatase activity. Triiodothyronine, a known stimulator of cartilage alkaline phosphatase, increased chick cartilage alkaline phosphatase activity 88% and phosphotyrosine phosphatase activity 106%, and stimulated porcine growth-plate cartilage alkaline phosphatase activity 91% and phosphotyrosine phosphatase activity 145% after 3 days of in vitro incubation. Each of the inhibitors block alkaline phosphatase and phosphotyrosine phosphatase activities. The purified alkaline phosphatase had a K
m for phosphotyrosine of 18 μmol/L and V
max of 5700 nmol tyrosine/mg protein/h, which is well over 1000-fold higher than the phosphotyrosine phosphatase activity found in the above preparations of pelvic and scapular cartilage. Furthermore, cartilage alkaline phosphatase dephosphorylated phosphotyrosinated histone and had little to no activity when histones phosphorylated at serine and threonine were used as substrate. Our studies support the hypothesis that alkaline phosphatase functions as a neutral phosphoprotein phosphatase, ie, a phosphotyrosine phosphatase, and is involved in the regulation of protein phosphorylation-dephosphorylation reactions.</description><subject>Alkaline Phosphatase - antagonists & inhibitors</subject><subject>Alkaline Phosphatase - metabolism</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cartilage - drug effects</subject><subject>Cartilage - embryology</subject><subject>Cartilage - enzymology</subject><subject>Chick Embryo</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Fetus - enzymology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Growth Plate - enzymology</subject><subject>Histocytochemistry</subject><subject>Histones - metabolism</subject><subject>Hydrogen-Ion Concentration</subject><subject>In Vitro Techniques</subject><subject>Kinetics</subject><subject>Phosphoprotein Phosphatases - antagonists & inhibitors</subject><subject>Phosphoprotein Phosphatases - metabolism</subject><subject>Phosphoric Monoester Hydrolases - antagonists & inhibitors</subject><subject>Phosphoric Monoester Hydrolases - metabolism</subject><subject>Skeleton and joints</subject><subject>Swine - embryology</subject><subject>Triiodothyronine - pharmacology</subject><subject>Vertebrates: osteoarticular system, musculoskeletal system</subject><issn>0026-0495</issn><issn>1532-8600</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1985</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1r3DAQhkVJSTZp_kEDPoSQHtyOPqyPSyCEfkGgPTRnMSvLiRKvvZG0ge2vr9w1S07tSWjeZ2Ze3iHkPYWPFKj8BMBkDcI0l7r5YIAyXfM3ZEEbzmotAQ7IYo8ckeOUHgFAKS0PySEzmoEyCxJ-Poxp_TDmbRxTGHyFQ1utd7V1HLMPw_zFjKnILoeXkLfV2FXYP2E_9bwGCr8qtViU32G4rxzGHHq89-_I2w775E_n94Tcffn86-Zbffvj6_eb69vaCc1z3SyFl1i8ceNRcJBgULbCsI4vpTLIDaAy0CDnHW1bJRoDxrWOURROeMpPyMVubrH_vPEp21VIzvc9Dn7cJKsaUxZR_V-QCiaVVhModqArGaXoO7uOYYVxaynY6RR2ytlOOVvd2L-nsLy0nc3zN8uVb_dNc_ZFP591TA77LuLgQtpjhilBuSjY1Q7zJbSX4KNNLvjB-TZE77Jtx_BvH38A6K2mrQ</recordid><startdate>198502</startdate><enddate>198502</enddate><creator>Burch, Warner M.</creator><creator>Hamner, Gail</creator><creator>Wuthier, Roy E.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7X8</scope></search><sort><creationdate>198502</creationdate><title>Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage</title><author>Burch, Warner M. ; Hamner, Gail ; Wuthier, Roy E.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c483t-5b4e6a07939ea430609a6d492f3b679a390a7905a33f1dd745909cdc21a4c4e13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1985</creationdate><topic>Alkaline Phosphatase - antagonists & inhibitors</topic><topic>Alkaline Phosphatase - metabolism</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cartilage - drug effects</topic><topic>Cartilage - embryology</topic><topic>Cartilage - enzymology</topic><topic>Chick Embryo</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Fetus - enzymology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Growth Plate - enzymology</topic><topic>Histocytochemistry</topic><topic>Histones - metabolism</topic><topic>Hydrogen-Ion Concentration</topic><topic>In Vitro Techniques</topic><topic>Kinetics</topic><topic>Phosphoprotein Phosphatases - antagonists & inhibitors</topic><topic>Phosphoprotein Phosphatases - metabolism</topic><topic>Phosphoric Monoester Hydrolases - antagonists & inhibitors</topic><topic>Phosphoric Monoester Hydrolases - metabolism</topic><topic>Skeleton and joints</topic><topic>Swine - embryology</topic><topic>Triiodothyronine - pharmacology</topic><topic>Vertebrates: osteoarticular system, musculoskeletal system</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Burch, Warner M.</creatorcontrib><creatorcontrib>Hamner, Gail</creatorcontrib><creatorcontrib>Wuthier, Roy E.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Metabolism, clinical and experimental</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Burch, Warner M.</au><au>Hamner, Gail</au><au>Wuthier, Roy E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage</atitle><jtitle>Metabolism, clinical and experimental</jtitle><addtitle>Metabolism</addtitle><date>1985-02</date><risdate>1985</risdate><volume>34</volume><issue>2</issue><spage>169</spage><epage>175</epage><pages>169-175</pages><issn>0026-0495</issn><eissn>1532-8600</eissn><abstract>We used embryonic skeletal cartilage known to have high levels of alkaline phosphatase activity to determine whether growing cartilage has phosphotyrosine phosphatase activity and phosphotyrosinyl histone phosphatase activity at physiologc pH. Embryonic chick pelvic cartilage and fetal pig scapular growth-plate cartilage were assayed using phosphotyrosine as substrate at pH 7.5 and the amount of tyrosine generated measured. Both cartilage models had K
m for phosphotyrosine between 6 to 24 μmol/L. Phosphotyrosine phosphatase activity correlated with alkaline phosphatase activity as assessed by (1) distribution of histologic staining for alkaline phosphatase within the cartilages, (2) hormonal stimulation of cartilage alkaline phosphatase activity in vitro, (3) comparison of alkaline phosphatase and phosphotyrosine phosphatase activities in the presence of known inhibitors (vanadate, levamisole, homoarginine, and zinc), and (4) assaying chick epiphyseal cartilage alkaline phosphatase purified to homogeneity for phosphotyrosine phosphatase activity. Areas of cartilage with elevated alkaline phosphatase activity also had raised phosphotyrosine phosphatase activity. Triiodothyronine, a known stimulator of cartilage alkaline phosphatase, increased chick cartilage alkaline phosphatase activity 88% and phosphotyrosine phosphatase activity 106%, and stimulated porcine growth-plate cartilage alkaline phosphatase activity 91% and phosphotyrosine phosphatase activity 145% after 3 days of in vitro incubation. Each of the inhibitors block alkaline phosphatase and phosphotyrosine phosphatase activities. The purified alkaline phosphatase had a K
m for phosphotyrosine of 18 μmol/L and V
max of 5700 nmol tyrosine/mg protein/h, which is well over 1000-fold higher than the phosphotyrosine phosphatase activity found in the above preparations of pelvic and scapular cartilage. Furthermore, cartilage alkaline phosphatase dephosphorylated phosphotyrosinated histone and had little to no activity when histones phosphorylated at serine and threonine were used as substrate. Our studies support the hypothesis that alkaline phosphatase functions as a neutral phosphoprotein phosphatase, ie, a phosphotyrosine phosphatase, and is involved in the regulation of protein phosphorylation-dephosphorylation reactions.</abstract><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>2982079</pmid><doi>10.1016/0026-0495(85)90128-3</doi><tpages>7</tpages></addata></record> |
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| ispartof | Metabolism, clinical and experimental, 1985-02, Vol.34 (2), p.169-175 |
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| subjects | Alkaline Phosphatase - antagonists & inhibitors Alkaline Phosphatase - metabolism Animals Biological and medical sciences Cartilage - drug effects Cartilage - embryology Cartilage - enzymology Chick Embryo Electrophoresis, Polyacrylamide Gel Fetus - enzymology Fundamental and applied biological sciences. Psychology Growth Plate - enzymology Histocytochemistry Histones - metabolism Hydrogen-Ion Concentration In Vitro Techniques Kinetics Phosphoprotein Phosphatases - antagonists & inhibitors Phosphoprotein Phosphatases - metabolism Phosphoric Monoester Hydrolases - antagonists & inhibitors Phosphoric Monoester Hydrolases - metabolism Skeleton and joints Swine - embryology Triiodothyronine - pharmacology Vertebrates: osteoarticular system, musculoskeletal system |
| title | Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage |
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