Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor
SaPIN2a encodes a proteinase inhibitor in nightshade ( Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized...
Gespeichert in:
| Veröffentlicht in: | Plant physiology and biochemistry 2007-10, Vol.45 (10), p.757-766 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 766 |
|---|---|
| container_issue | 10 |
| container_start_page | 757 |
| container_title | Plant physiology and biochemistry |
| container_volume | 45 |
| creator | Wang, Zhen-Yu Ding, Ling-Wen Ge, Zhi-Juan Wang, Zhaoyu Wang, Fanghai Li, Ning Xu, Zeng-Fu |
| description | SaPIN2a encodes a proteinase inhibitor in nightshade (
Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized native SaPIN2a from nightshade stems and recombinant SaPIN2a expressed in
Escherichia coli. Purified native SaPIN2a was found as a charge isomer family of homodimers, and was weakly glycosylated. Native SaPIN2a significantly inhibited serine proteinases such as trypsin, chymotrypsin, and subtilisin, with the most potent inhibitory activity on subtilisin. It did not inhibit cysteine proteinase papain and aspartic proteinase cathepsin D. Recombinant SaPIN2a had a strong inhibitory effect on chymotrypsin, but its inhibitory activities toward trypsin and especially toward subtilisin were greatly reduced. In addition, native SaPIN2a can effectively inhibit midgut trypsin-like activities from
Trichoplusia ni and
Spodoptera litura larvae, suggesting a potential for the production of insect-resistant transgenic plants. |
| doi_str_mv | 10.1016/j.plaphy.2007.07.012 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_68397231</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0981942807001532</els_id><sourcerecordid>19807962</sourcerecordid><originalsourceid>FETCH-LOGICAL-c421t-f6cc34dea177d9af46bc0e4bbe01c23fa8df9872d8556b904f65e857066bd05c3</originalsourceid><addsrcrecordid>eNqFkVuLFDEQhYMo7rj6D0T6RZ_sMUmnc3kRlsXLwqIL6nNIJxUmQ3dnTDILs7_etD2wbwoFCVXfKQ51EHpN8JZgwj_st4fRHHanLcVYbJci9AnaECm6lnKFn6INVpK0ilF5gV7kvMcYUya65-iCCClwr-gGne6OKfhgTQlxbszsGrszydgCKTyszeibuf7u4e84gY3TEGYzl-aHubv5Rs37xjTVS23kABWDESaYSztGa8bwAK45pFigajI0Yd6FIZSYXqJn3owZXp3fS_Tr86ef11_b2-9fbq6vblvLKCmt59Z2zIEhQjhlPOODxcCGATCxtPNGOq-koE72PR8UZp73IHuBOR8c7m13id6te6uJ30fIRU8hWxirYYjHrLnslKAd-S9IlMRCcVpBtoI2xZwTeH1IYTLppAnWSzZ6r9ds9JKNXoossjfn_cdhAvcoOodRgbdnwOR6OZ_MbEN-5BQRTGFRuY8rB_Vs9wGSzjbAbMGFmk7RLoZ_O_kD-4SxPQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>19807962</pqid></control><display><type>article</type><title>Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Wang, Zhen-Yu ; Ding, Ling-Wen ; Ge, Zhi-Juan ; Wang, Zhaoyu ; Wang, Fanghai ; Li, Ning ; Xu, Zeng-Fu</creator><creatorcontrib>Wang, Zhen-Yu ; Ding, Ling-Wen ; Ge, Zhi-Juan ; Wang, Zhaoyu ; Wang, Fanghai ; Li, Ning ; Xu, Zeng-Fu</creatorcontrib><description>SaPIN2a encodes a proteinase inhibitor in nightshade (
Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized native SaPIN2a from nightshade stems and recombinant SaPIN2a expressed in
Escherichia coli. Purified native SaPIN2a was found as a charge isomer family of homodimers, and was weakly glycosylated. Native SaPIN2a significantly inhibited serine proteinases such as trypsin, chymotrypsin, and subtilisin, with the most potent inhibitory activity on subtilisin. It did not inhibit cysteine proteinase papain and aspartic proteinase cathepsin D. Recombinant SaPIN2a had a strong inhibitory effect on chymotrypsin, but its inhibitory activities toward trypsin and especially toward subtilisin were greatly reduced. In addition, native SaPIN2a can effectively inhibit midgut trypsin-like activities from
Trichoplusia ni and
Spodoptera litura larvae, suggesting a potential for the production of insect-resistant transgenic plants.</description><identifier>ISSN: 0981-9428</identifier><identifier>EISSN: 1873-2690</identifier><identifier>DOI: 10.1016/j.plaphy.2007.07.012</identifier><identifier>PMID: 17870592</identifier><identifier>CODEN: PPBIEX</identifier><language>eng</language><publisher>Paris: Elsevier Masson SAS</publisher><subject>Amino Acid Sequence ; Biological and medical sciences ; Chymotrypsin - antagonists & inhibitors ; Chymotrypsin - metabolism ; Cysteine Endopeptidases - metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzymes ; Escherichia coli ; Fundamental and applied biological sciences. Psychology ; Hydrogen-Ion Concentration ; Insect ; Kinetics ; Metabolism ; Molecular Sequence Data ; Phloem ; Phloem - genetics ; Phloem - metabolism ; Plant physiology and development ; Plant Proteins - genetics ; Plant Proteins - isolation & purification ; Plant Proteins - metabolism ; Plant Stems - genetics ; Plant Stems - metabolism ; Protease inhibitor ; Protease Inhibitors - chemistry ; Protease Inhibitors - metabolism ; Protease Inhibitors - pharmacology ; Proteinase ; Proteolysis ; Recombinant Proteins - metabolism ; Recombinant Proteins - pharmacology ; Sequence Analysis, Protein ; Serine Endopeptidases - metabolism ; Sieve element ; Solanum - genetics ; Solanum - metabolism ; Solanum americanum ; Spodoptera litura ; Subtilisin - antagonists & inhibitors ; Subtilisin - metabolism ; Temperature ; Trichoplusia ni ; Trypsin - metabolism</subject><ispartof>Plant physiology and biochemistry, 2007-10, Vol.45 (10), p.757-766</ispartof><rights>2007</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c421t-f6cc34dea177d9af46bc0e4bbe01c23fa8df9872d8556b904f65e857066bd05c3</citedby><cites>FETCH-LOGICAL-c421t-f6cc34dea177d9af46bc0e4bbe01c23fa8df9872d8556b904f65e857066bd05c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0981942807001532$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19174907$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17870592$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Zhen-Yu</creatorcontrib><creatorcontrib>Ding, Ling-Wen</creatorcontrib><creatorcontrib>Ge, Zhi-Juan</creatorcontrib><creatorcontrib>Wang, Zhaoyu</creatorcontrib><creatorcontrib>Wang, Fanghai</creatorcontrib><creatorcontrib>Li, Ning</creatorcontrib><creatorcontrib>Xu, Zeng-Fu</creatorcontrib><title>Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor</title><title>Plant physiology and biochemistry</title><addtitle>Plant Physiol Biochem</addtitle><description>SaPIN2a encodes a proteinase inhibitor in nightshade (
Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized native SaPIN2a from nightshade stems and recombinant SaPIN2a expressed in
Escherichia coli. Purified native SaPIN2a was found as a charge isomer family of homodimers, and was weakly glycosylated. Native SaPIN2a significantly inhibited serine proteinases such as trypsin, chymotrypsin, and subtilisin, with the most potent inhibitory activity on subtilisin. It did not inhibit cysteine proteinase papain and aspartic proteinase cathepsin D. Recombinant SaPIN2a had a strong inhibitory effect on chymotrypsin, but its inhibitory activities toward trypsin and especially toward subtilisin were greatly reduced. In addition, native SaPIN2a can effectively inhibit midgut trypsin-like activities from
Trichoplusia ni and
Spodoptera litura larvae, suggesting a potential for the production of insect-resistant transgenic plants.</description><subject>Amino Acid Sequence</subject><subject>Biological and medical sciences</subject><subject>Chymotrypsin - antagonists & inhibitors</subject><subject>Chymotrypsin - metabolism</subject><subject>Cysteine Endopeptidases - metabolism</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzymes</subject><subject>Escherichia coli</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hydrogen-Ion Concentration</subject><subject>Insect</subject><subject>Kinetics</subject><subject>Metabolism</subject><subject>Molecular Sequence Data</subject><subject>Phloem</subject><subject>Phloem - genetics</subject><subject>Phloem - metabolism</subject><subject>Plant physiology and development</subject><subject>Plant Proteins - genetics</subject><subject>Plant Proteins - isolation & purification</subject><subject>Plant Proteins - metabolism</subject><subject>Plant Stems - genetics</subject><subject>Plant Stems - metabolism</subject><subject>Protease inhibitor</subject><subject>Protease Inhibitors - chemistry</subject><subject>Protease Inhibitors - metabolism</subject><subject>Protease Inhibitors - pharmacology</subject><subject>Proteinase</subject><subject>Proteolysis</subject><subject>Recombinant Proteins - metabolism</subject><subject>Recombinant Proteins - pharmacology</subject><subject>Sequence Analysis, Protein</subject><subject>Serine Endopeptidases - metabolism</subject><subject>Sieve element</subject><subject>Solanum - genetics</subject><subject>Solanum - metabolism</subject><subject>Solanum americanum</subject><subject>Spodoptera litura</subject><subject>Subtilisin - antagonists & inhibitors</subject><subject>Subtilisin - metabolism</subject><subject>Temperature</subject><subject>Trichoplusia ni</subject><subject>Trypsin - metabolism</subject><issn>0981-9428</issn><issn>1873-2690</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkVuLFDEQhYMo7rj6D0T6RZ_sMUmnc3kRlsXLwqIL6nNIJxUmQ3dnTDILs7_etD2wbwoFCVXfKQ51EHpN8JZgwj_st4fRHHanLcVYbJci9AnaECm6lnKFn6INVpK0ilF5gV7kvMcYUya65-iCCClwr-gGne6OKfhgTQlxbszsGrszydgCKTyszeibuf7u4e84gY3TEGYzl-aHubv5Rs37xjTVS23kABWDESaYSztGa8bwAK45pFigajI0Yd6FIZSYXqJn3owZXp3fS_Tr86ef11_b2-9fbq6vblvLKCmt59Z2zIEhQjhlPOODxcCGATCxtPNGOq-koE72PR8UZp73IHuBOR8c7m13id6te6uJ30fIRU8hWxirYYjHrLnslKAd-S9IlMRCcVpBtoI2xZwTeH1IYTLppAnWSzZ6r9ds9JKNXoossjfn_cdhAvcoOodRgbdnwOR6OZ_MbEN-5BQRTGFRuY8rB_Vs9wGSzjbAbMGFmk7RLoZ_O_kD-4SxPQ</recordid><startdate>20071001</startdate><enddate>20071001</enddate><creator>Wang, Zhen-Yu</creator><creator>Ding, Ling-Wen</creator><creator>Ge, Zhi-Juan</creator><creator>Wang, Zhaoyu</creator><creator>Wang, Fanghai</creator><creator>Li, Ning</creator><creator>Xu, Zeng-Fu</creator><general>Elsevier Masson SAS</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>7QL</scope><scope>7SS</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>20071001</creationdate><title>Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor</title><author>Wang, Zhen-Yu ; Ding, Ling-Wen ; Ge, Zhi-Juan ; Wang, Zhaoyu ; Wang, Fanghai ; Li, Ning ; Xu, Zeng-Fu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c421t-f6cc34dea177d9af46bc0e4bbe01c23fa8df9872d8556b904f65e857066bd05c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Amino Acid Sequence</topic><topic>Biological and medical sciences</topic><topic>Chymotrypsin - antagonists & inhibitors</topic><topic>Chymotrypsin - metabolism</topic><topic>Cysteine Endopeptidases - metabolism</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzymes</topic><topic>Escherichia coli</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hydrogen-Ion Concentration</topic><topic>Insect</topic><topic>Kinetics</topic><topic>Metabolism</topic><topic>Molecular Sequence Data</topic><topic>Phloem</topic><topic>Phloem - genetics</topic><topic>Phloem - metabolism</topic><topic>Plant physiology and development</topic><topic>Plant Proteins - genetics</topic><topic>Plant Proteins - isolation & purification</topic><topic>Plant Proteins - metabolism</topic><topic>Plant Stems - genetics</topic><topic>Plant Stems - metabolism</topic><topic>Protease inhibitor</topic><topic>Protease Inhibitors - chemistry</topic><topic>Protease Inhibitors - metabolism</topic><topic>Protease Inhibitors - pharmacology</topic><topic>Proteinase</topic><topic>Proteolysis</topic><topic>Recombinant Proteins - metabolism</topic><topic>Recombinant Proteins - pharmacology</topic><topic>Sequence Analysis, Protein</topic><topic>Serine Endopeptidases - metabolism</topic><topic>Sieve element</topic><topic>Solanum - genetics</topic><topic>Solanum - metabolism</topic><topic>Solanum americanum</topic><topic>Spodoptera litura</topic><topic>Subtilisin - antagonists & inhibitors</topic><topic>Subtilisin - metabolism</topic><topic>Temperature</topic><topic>Trichoplusia ni</topic><topic>Trypsin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Zhen-Yu</creatorcontrib><creatorcontrib>Ding, Ling-Wen</creatorcontrib><creatorcontrib>Ge, Zhi-Juan</creatorcontrib><creatorcontrib>Wang, Zhaoyu</creatorcontrib><creatorcontrib>Wang, Fanghai</creatorcontrib><creatorcontrib>Li, Ning</creatorcontrib><creatorcontrib>Xu, Zeng-Fu</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Plant physiology and biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Zhen-Yu</au><au>Ding, Ling-Wen</au><au>Ge, Zhi-Juan</au><au>Wang, Zhaoyu</au><au>Wang, Fanghai</au><au>Li, Ning</au><au>Xu, Zeng-Fu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor</atitle><jtitle>Plant physiology and biochemistry</jtitle><addtitle>Plant Physiol Biochem</addtitle><date>2007-10-01</date><risdate>2007</risdate><volume>45</volume><issue>10</issue><spage>757</spage><epage>766</epage><pages>757-766</pages><issn>0981-9428</issn><eissn>1873-2690</eissn><coden>PPBIEX</coden><abstract>SaPIN2a encodes a proteinase inhibitor in nightshade (
Solanum americanum), which is specifically localized to the enucleate sieve elements. It has been proposed to play an important role in phloem development by regulating proteolysis in sieve elements. In this study, we purified and characterized native SaPIN2a from nightshade stems and recombinant SaPIN2a expressed in
Escherichia coli. Purified native SaPIN2a was found as a charge isomer family of homodimers, and was weakly glycosylated. Native SaPIN2a significantly inhibited serine proteinases such as trypsin, chymotrypsin, and subtilisin, with the most potent inhibitory activity on subtilisin. It did not inhibit cysteine proteinase papain and aspartic proteinase cathepsin D. Recombinant SaPIN2a had a strong inhibitory effect on chymotrypsin, but its inhibitory activities toward trypsin and especially toward subtilisin were greatly reduced. In addition, native SaPIN2a can effectively inhibit midgut trypsin-like activities from
Trichoplusia ni and
Spodoptera litura larvae, suggesting a potential for the production of insect-resistant transgenic plants.</abstract><cop>Paris</cop><pub>Elsevier Masson SAS</pub><pmid>17870592</pmid><doi>10.1016/j.plaphy.2007.07.012</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0981-9428 |
| ispartof | Plant physiology and biochemistry, 2007-10, Vol.45 (10), p.757-766 |
| issn | 0981-9428 1873-2690 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_68397231 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Amino Acid Sequence Biological and medical sciences Chymotrypsin - antagonists & inhibitors Chymotrypsin - metabolism Cysteine Endopeptidases - metabolism Electrophoresis, Polyacrylamide Gel Enzymes Escherichia coli Fundamental and applied biological sciences. Psychology Hydrogen-Ion Concentration Insect Kinetics Metabolism Molecular Sequence Data Phloem Phloem - genetics Phloem - metabolism Plant physiology and development Plant Proteins - genetics Plant Proteins - isolation & purification Plant Proteins - metabolism Plant Stems - genetics Plant Stems - metabolism Protease inhibitor Protease Inhibitors - chemistry Protease Inhibitors - metabolism Protease Inhibitors - pharmacology Proteinase Proteolysis Recombinant Proteins - metabolism Recombinant Proteins - pharmacology Sequence Analysis, Protein Serine Endopeptidases - metabolism Sieve element Solanum - genetics Solanum - metabolism Solanum americanum Spodoptera litura Subtilisin - antagonists & inhibitors Subtilisin - metabolism Temperature Trichoplusia ni Trypsin - metabolism |
| title | Purification and characterization of native and recombinant SaPIN2a, a plant sieve element-localized proteinase inhibitor |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-16T04%3A00%3A02IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Purification%20and%20characterization%20of%20native%20and%20recombinant%20SaPIN2a,%20a%20plant%20sieve%20element-localized%20proteinase%20inhibitor&rft.jtitle=Plant%20physiology%20and%20biochemistry&rft.au=Wang,%20Zhen-Yu&rft.date=2007-10-01&rft.volume=45&rft.issue=10&rft.spage=757&rft.epage=766&rft.pages=757-766&rft.issn=0981-9428&rft.eissn=1873-2690&rft.coden=PPBIEX&rft_id=info:doi/10.1016/j.plaphy.2007.07.012&rft_dat=%3Cproquest_cross%3E19807962%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=19807962&rft_id=info:pmid/17870592&rft_els_id=S0981942807001532&rfr_iscdi=true |