A new purification procedure for bovine milk xanthine oxidase: Effect of proteolysis on the subunit structure
A new purification procedure for bovine milk xanthine oxidase is reported. The enzyme so obtained is of the highest purity and shows little evidence of degradation. The enzyme displays a single protein band on either polyacrylamide gels or on sodium dodecyl sulfate-urea polyacrylamide gels. Sediment...
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| Veröffentlicht in: | Archives of biochemistry and biophysics 1975-01, Vol.169 (2), p.695-701 |
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| container_end_page | 701 |
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| container_issue | 2 |
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| container_title | Archives of biochemistry and biophysics |
| container_volume | 169 |
| creator | Waud, William R. Brady, Frank O. Wiley, Ralph D. Rajagopalan, K.V. |
| description | A new purification procedure for bovine milk xanthine oxidase is reported. The enzyme so obtained is of the highest purity and shows little evidence of degradation. The enzyme displays a single protein band on either polyacrylamide gels or on sodium dodecyl sulfate-urea polyacrylamide gels. Sedimentation equilibrium studies indicate a native molecular weight of 303,000 and a subunit molecular weight of approximately 150,000. The latter value is in good agreement with the minimum molecular weight of 157,000 calculated from dry weight determination and flavin analysis. In contrast, purification of xanthine oxidase from pancreatin-treated cream yields a protein which displays two subunits corresponding to molecular weights of 92,000 and 39,000 as determined by dodecyl sulfate-urea polyacrylamide gel electrophoresis. Pancreatinized enzyme has a greater mobility than unproteolyzed enzyme on polyacrylamide gels. Exposure of milk xanthine oxidase to pancreatin before isolation or after purification yields the same result. |
| doi_str_mv | 10.1016/0003-9861(75)90214-3 |
| format | Article |
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The enzyme so obtained is of the highest purity and shows little evidence of degradation. The enzyme displays a single protein band on either polyacrylamide gels or on sodium dodecyl sulfate-urea polyacrylamide gels. Sedimentation equilibrium studies indicate a native molecular weight of 303,000 and a subunit molecular weight of approximately 150,000. The latter value is in good agreement with the minimum molecular weight of 157,000 calculated from dry weight determination and flavin analysis. In contrast, purification of xanthine oxidase from pancreatin-treated cream yields a protein which displays two subunits corresponding to molecular weights of 92,000 and 39,000 as determined by dodecyl sulfate-urea polyacrylamide gel electrophoresis. Pancreatinized enzyme has a greater mobility than unproteolyzed enzyme on polyacrylamide gels. Exposure of milk xanthine oxidase to pancreatin before isolation or after purification yields the same result.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1016/0003-9861(75)90214-3</identifier><identifier>PMID: 1180567</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>animal science ; Animals ; Cattle ; Chromatography, Ion Exchange ; Electrophoresis, Disc ; Electrophoresis, Polyacrylamide Gel ; Female ; Flavins - analysis ; livestock ; Macromolecular Substances ; Methods ; Milk - enzymology ; Molecular Weight ; Pancreatin ; Spectrophotometry ; Spectrophotometry, Ultraviolet ; Xanthine Oxidase - isolation & purification ; Xanthine Oxidase - metabolism ; zoology</subject><ispartof>Archives of biochemistry and biophysics, 1975-01, Vol.169 (2), p.695-701</ispartof><rights>1975</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c381t-2bebe911d9b65123f6db4b9938ac46194511a0d9cf54504f9ef2731bbc00ecb43</citedby><cites>FETCH-LOGICAL-c381t-2bebe911d9b65123f6db4b9938ac46194511a0d9cf54504f9ef2731bbc00ecb43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-9861(75)90214-3$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1180567$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Waud, William R.</creatorcontrib><creatorcontrib>Brady, Frank O.</creatorcontrib><creatorcontrib>Wiley, Ralph D.</creatorcontrib><creatorcontrib>Rajagopalan, K.V.</creatorcontrib><title>A new purification procedure for bovine milk xanthine oxidase: Effect of proteolysis on the subunit structure</title><title>Archives of biochemistry and biophysics</title><addtitle>Arch Biochem Biophys</addtitle><description>A new purification procedure for bovine milk xanthine oxidase is reported. The enzyme so obtained is of the highest purity and shows little evidence of degradation. The enzyme displays a single protein band on either polyacrylamide gels or on sodium dodecyl sulfate-urea polyacrylamide gels. Sedimentation equilibrium studies indicate a native molecular weight of 303,000 and a subunit molecular weight of approximately 150,000. The latter value is in good agreement with the minimum molecular weight of 157,000 calculated from dry weight determination and flavin analysis. In contrast, purification of xanthine oxidase from pancreatin-treated cream yields a protein which displays two subunits corresponding to molecular weights of 92,000 and 39,000 as determined by dodecyl sulfate-urea polyacrylamide gel electrophoresis. Pancreatinized enzyme has a greater mobility than unproteolyzed enzyme on polyacrylamide gels. Exposure of milk xanthine oxidase to pancreatin before isolation or after purification yields the same result.</description><subject>animal science</subject><subject>Animals</subject><subject>Cattle</subject><subject>Chromatography, Ion Exchange</subject><subject>Electrophoresis, Disc</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Female</subject><subject>Flavins - analysis</subject><subject>livestock</subject><subject>Macromolecular Substances</subject><subject>Methods</subject><subject>Milk - enzymology</subject><subject>Molecular Weight</subject><subject>Pancreatin</subject><subject>Spectrophotometry</subject><subject>Spectrophotometry, Ultraviolet</subject><subject>Xanthine Oxidase - isolation & purification</subject><subject>Xanthine Oxidase - metabolism</subject><subject>zoology</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1975</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtu1TAQhi0EKocDbwDCKwSLgCe2k5gFUlWVi1SJBXRt2c6YGk7ig-2U9u1xSAU7VqPRf9Gvj5CnwF4Dg-4NY4w3aujgZS9fKdaCaPg9sgOmuobxQdwnu7-Wh-RRzt8ZAxBde0JOAAYmu35HplM64y96XFLwwZkS4kyPKTocl4TUx0RtvA4z0ikcftAbM5er9Ys3YTQZ39Jz79EVGv2aKhgPtzlkWkvKFdK82GUOheaSFldq4WPywJtDxid3d08u359_PfvYXHz-8Ons9KJxfIDStBYtKoBR2U5Cy303WmGV4oNxogMlJIBho3JeCsmEV-jbnoO1jjF0VvA9ebH11lE_F8xFTyE7PBzMjHHJeuBMSFnh7InYjC7FnBN6fUxhMulWA9MrZb0i1CtC3Uv9h7LmNfbsrn-xE47_QhvWqj_fdG-iNt9SyPryS8uAs7av-2VbHe82B1YM1wGTzi7gXLmHVIHqMYb_T_gNElOWew</recordid><startdate>19750101</startdate><enddate>19750101</enddate><creator>Waud, William R.</creator><creator>Brady, Frank O.</creator><creator>Wiley, Ralph D.</creator><creator>Rajagopalan, K.V.</creator><general>Elsevier Inc</general><scope>FBQ</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>7X8</scope></search><sort><creationdate>19750101</creationdate><title>A new purification procedure for bovine milk xanthine oxidase: Effect of proteolysis on the subunit structure</title><author>Waud, William R. ; Brady, Frank O. ; Wiley, Ralph D. ; Rajagopalan, K.V.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c381t-2bebe911d9b65123f6db4b9938ac46194511a0d9cf54504f9ef2731bbc00ecb43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1975</creationdate><topic>animal science</topic><topic>Animals</topic><topic>Cattle</topic><topic>Chromatography, Ion Exchange</topic><topic>Electrophoresis, Disc</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Female</topic><topic>Flavins - analysis</topic><topic>livestock</topic><topic>Macromolecular Substances</topic><topic>Methods</topic><topic>Milk - enzymology</topic><topic>Molecular Weight</topic><topic>Pancreatin</topic><topic>Spectrophotometry</topic><topic>Spectrophotometry, Ultraviolet</topic><topic>Xanthine Oxidase - isolation & purification</topic><topic>Xanthine Oxidase - metabolism</topic><topic>zoology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Waud, William R.</creatorcontrib><creatorcontrib>Brady, Frank O.</creatorcontrib><creatorcontrib>Wiley, Ralph D.</creatorcontrib><creatorcontrib>Rajagopalan, K.V.</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Archives of biochemistry and biophysics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Waud, William R.</au><au>Brady, Frank O.</au><au>Wiley, Ralph D.</au><au>Rajagopalan, K.V.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A new purification procedure for bovine milk xanthine oxidase: Effect of proteolysis on the subunit structure</atitle><jtitle>Archives of biochemistry and biophysics</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>1975-01-01</date><risdate>1975</risdate><volume>169</volume><issue>2</issue><spage>695</spage><epage>701</epage><pages>695-701</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><abstract>A new purification procedure for bovine milk xanthine oxidase is reported. The enzyme so obtained is of the highest purity and shows little evidence of degradation. The enzyme displays a single protein band on either polyacrylamide gels or on sodium dodecyl sulfate-urea polyacrylamide gels. Sedimentation equilibrium studies indicate a native molecular weight of 303,000 and a subunit molecular weight of approximately 150,000. The latter value is in good agreement with the minimum molecular weight of 157,000 calculated from dry weight determination and flavin analysis. In contrast, purification of xanthine oxidase from pancreatin-treated cream yields a protein which displays two subunits corresponding to molecular weights of 92,000 and 39,000 as determined by dodecyl sulfate-urea polyacrylamide gel electrophoresis. Pancreatinized enzyme has a greater mobility than unproteolyzed enzyme on polyacrylamide gels. Exposure of milk xanthine oxidase to pancreatin before isolation or after purification yields the same result.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>1180567</pmid><doi>10.1016/0003-9861(75)90214-3</doi><tpages>7</tpages></addata></record> |
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| identifier | ISSN: 0003-9861 |
| ispartof | Archives of biochemistry and biophysics, 1975-01, Vol.169 (2), p.695-701 |
| issn | 0003-9861 1096-0384 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | animal science Animals Cattle Chromatography, Ion Exchange Electrophoresis, Disc Electrophoresis, Polyacrylamide Gel Female Flavins - analysis livestock Macromolecular Substances Methods Milk - enzymology Molecular Weight Pancreatin Spectrophotometry Spectrophotometry, Ultraviolet Xanthine Oxidase - isolation & purification Xanthine Oxidase - metabolism zoology |
| title | A new purification procedure for bovine milk xanthine oxidase: Effect of proteolysis on the subunit structure |
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