Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells
The maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. The putative processing enzyme, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (r...
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| Veröffentlicht in: | Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2011, Vol.75 (12), p.2371-2375 |
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| container_title | Bioscience, biotechnology, and biochemistry |
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| creator | HSIEH, Sheng-Kuo TZEN, Jason T. C. WU, Tzong-Yuan CHEN, Ying-Ju YANG, Wei-Hung HUANG, Chun-Fa HSIEH, Feng-Chia JINN, Tzyy-Rong |
| description | The maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. The putative processing enzyme, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (rDPP-IV) via the baculovirus expression system. rDPP-IV purified by one-step nickel-affinity chromatography was verified by Western blot and LC-MS/MS analysis. The k
cat
/K
m
of rDPP-IV was determined to be in the range of 10-500 mM
−1
·S
−1
for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50 °C and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80 and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride respectively. |
| doi_str_mv | 10.1271/bbb.110571 |
| format | Article |
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cat
/K
m
of rDPP-IV was determined to be in the range of 10-500 mM
−1
·S
−1
for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50 °C and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80 and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride respectively.</description><identifier>ISSN: 0916-8451</identifier><identifier>EISSN: 1347-6947</identifier><identifier>DOI: 10.1271/bbb.110571</identifier><identifier>PMID: 22146730</identifier><language>eng</language><publisher>Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry</publisher><subject>Amino Acid Sequence ; Animals ; Baculoviridae - genetics ; Biological and medical sciences ; Cell Line ; Chromatography, Affinity ; Dipeptidyl Peptidase 4 - chemistry ; Dipeptidyl Peptidase 4 - genetics ; Dipeptidyl Peptidase 4 - isolation & purification ; Dipeptidyl Peptidase 4 - metabolism ; dipeptidyl peptidase IV ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; mastoparan B ; Molecular Sequence Data ; prosequence processing ; sitagliptin ; Vespa basalis ; Wasps - enzymology ; Wasps - genetics</subject><ispartof>Bioscience, biotechnology, and biochemistry, 2011, Vol.75 (12), p.2371-2375</ispartof><rights>2011 by Japan Society for Bioscience, Biotechnology, and Agrochemistry 2011</rights><rights>2015 INIST-CNRS</rights><rights>Copyright Japan Science and Technology Agency 2011</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c518t-f5d848b95cd2c81fc5f3efdbab453b259667a480ee83d8db074e222fd571002c3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,778,782,4012,27910,27911,27912</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=25615477$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22146730$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>HSIEH, Sheng-Kuo</creatorcontrib><creatorcontrib>TZEN, Jason T. C.</creatorcontrib><creatorcontrib>WU, Tzong-Yuan</creatorcontrib><creatorcontrib>CHEN, Ying-Ju</creatorcontrib><creatorcontrib>YANG, Wei-Hung</creatorcontrib><creatorcontrib>HUANG, Chun-Fa</creatorcontrib><creatorcontrib>HSIEH, Feng-Chia</creatorcontrib><creatorcontrib>JINN, Tzyy-Rong</creatorcontrib><title>Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells</title><title>Bioscience, biotechnology, and biochemistry</title><addtitle>Biosci Biotechnol Biochem</addtitle><description>The maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. The putative processing enzyme, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (rDPP-IV) via the baculovirus expression system. rDPP-IV purified by one-step nickel-affinity chromatography was verified by Western blot and LC-MS/MS analysis. The k
cat
/K
m
of rDPP-IV was determined to be in the range of 10-500 mM
−1
·S
−1
for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50 °C and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80 and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride respectively.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Baculoviridae - genetics</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Chromatography, Affinity</subject><subject>Dipeptidyl Peptidase 4 - chemistry</subject><subject>Dipeptidyl Peptidase 4 - genetics</subject><subject>Dipeptidyl Peptidase 4 - isolation & purification</subject><subject>Dipeptidyl Peptidase 4 - metabolism</subject><subject>dipeptidyl peptidase IV</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>mastoparan B</subject><subject>Molecular Sequence Data</subject><subject>prosequence processing</subject><subject>sitagliptin</subject><subject>Vespa basalis</subject><subject>Wasps - enzymology</subject><subject>Wasps - genetics</subject><issn>0916-8451</issn><issn>1347-6947</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpt0V1rFDEUBuAgil2rN_4ACYgIwtQkk8zHpV1bu1BQUHs7nHzR1OxkzJlB1x_g7zZ1twriVT54Eg7vS8hTzk64aPlrrfUJ50y1_B5Z8Vq2VdPL9j5ZsZ43VScVPyKPEG8YKxeKPyRHQnDZtDVbkZ_ny2jmkEaI9Oz7lB1iOVAYLV1fQwYzuxx-wK2gydO3YXLTHOwu0g-_N4CObq6oz2lL52tHTyOYL9WpizE4Sy9SHt1MrxxOQDUgxIA0jPSjF5xuRnRmputi8TF54CGie3JYj8nn87NP64vq8v27zfrNZWUU7-bKK9vJTvfKWGE67o3ytfNWg5aq1kL1TdOC7JhzXW07q1krnRDC25INY8LUx-Tl_t8pp6-Lw3nYBjRlAhhdWnDoS6C9qmVf5PN_5E1acokJBy5lX1Jloivq1V6ZnBCz88OUwxbybuBsuC1nKOUM-3IKfnb4ctFbZ__QuzYKeHEAgAaizzCagH-dariSbVuc2rsw-pS38C3laIcZdjHlu0f1fwb4BcaVqUI</recordid><startdate>2011</startdate><enddate>2011</enddate><creator>HSIEH, Sheng-Kuo</creator><creator>TZEN, Jason T. C.</creator><creator>WU, Tzong-Yuan</creator><creator>CHEN, Ying-Ju</creator><creator>YANG, Wei-Hung</creator><creator>HUANG, Chun-Fa</creator><creator>HSIEH, Feng-Chia</creator><creator>JINN, Tzyy-Rong</creator><general>Japan Society for Bioscience, Biotechnology, and Agrochemistry</general><general>Japan Society for Bioscience Biotechnology and Agrochemistry</general><general>Oxford University Press</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>7X8</scope></search><sort><creationdate>2011</creationdate><title>Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells</title><author>HSIEH, Sheng-Kuo ; TZEN, Jason T. C. ; WU, Tzong-Yuan ; CHEN, Ying-Ju ; YANG, Wei-Hung ; HUANG, Chun-Fa ; HSIEH, Feng-Chia ; JINN, Tzyy-Rong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c518t-f5d848b95cd2c81fc5f3efdbab453b259667a480ee83d8db074e222fd571002c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Baculoviridae - genetics</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Chromatography, Affinity</topic><topic>Dipeptidyl Peptidase 4 - chemistry</topic><topic>Dipeptidyl Peptidase 4 - genetics</topic><topic>Dipeptidyl Peptidase 4 - isolation & purification</topic><topic>Dipeptidyl Peptidase 4 - metabolism</topic><topic>dipeptidyl peptidase IV</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>mastoparan B</topic><topic>Molecular Sequence Data</topic><topic>prosequence processing</topic><topic>sitagliptin</topic><topic>Vespa basalis</topic><topic>Wasps - enzymology</topic><topic>Wasps - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>HSIEH, Sheng-Kuo</creatorcontrib><creatorcontrib>TZEN, Jason T. C.</creatorcontrib><creatorcontrib>WU, Tzong-Yuan</creatorcontrib><creatorcontrib>CHEN, Ying-Ju</creatorcontrib><creatorcontrib>YANG, Wei-Hung</creatorcontrib><creatorcontrib>HUANG, Chun-Fa</creatorcontrib><creatorcontrib>HSIEH, Feng-Chia</creatorcontrib><creatorcontrib>JINN, Tzyy-Rong</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>MEDLINE - Academic</collection><jtitle>Bioscience, biotechnology, and biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>HSIEH, Sheng-Kuo</au><au>TZEN, Jason T. C.</au><au>WU, Tzong-Yuan</au><au>CHEN, Ying-Ju</au><au>YANG, Wei-Hung</au><au>HUANG, Chun-Fa</au><au>HSIEH, Feng-Chia</au><au>JINN, Tzyy-Rong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells</atitle><jtitle>Bioscience, biotechnology, and biochemistry</jtitle><addtitle>Biosci Biotechnol Biochem</addtitle><date>2011</date><risdate>2011</risdate><volume>75</volume><issue>12</issue><spage>2371</spage><epage>2375</epage><pages>2371-2375</pages><issn>0916-8451</issn><eissn>1347-6947</eissn><abstract>The maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. The putative processing enzyme, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (rDPP-IV) via the baculovirus expression system. rDPP-IV purified by one-step nickel-affinity chromatography was verified by Western blot and LC-MS/MS analysis. The k
cat
/K
m
of rDPP-IV was determined to be in the range of 10-500 mM
−1
·S
−1
for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50 °C and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80 and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride respectively.</abstract><cop>Tokyo</cop><pub>Japan Society for Bioscience, Biotechnology, and Agrochemistry</pub><pmid>22146730</pmid><doi>10.1271/bbb.110571</doi><tpages>5</tpages></addata></record> |
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| ispartof | Bioscience, biotechnology, and biochemistry, 2011, Vol.75 (12), p.2371-2375 |
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| source | J-STAGE Free; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Oxford University Press Journals All Titles (1996-Current); Freely Accessible Japanese Titles; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence Animals Baculoviridae - genetics Biological and medical sciences Cell Line Chromatography, Affinity Dipeptidyl Peptidase 4 - chemistry Dipeptidyl Peptidase 4 - genetics Dipeptidyl Peptidase 4 - isolation & purification Dipeptidyl Peptidase 4 - metabolism dipeptidyl peptidase IV Fundamental and applied biological sciences. Psychology Gene Expression mastoparan B Molecular Sequence Data prosequence processing sitagliptin Vespa basalis Wasps - enzymology Wasps - genetics |
| title | Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells |
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