Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom
Leucurolysin-B (leuc-B) is an hemorrhagic metalloproteinase found in the venom of Bothrops leucurus (white-tailed-jararaca) snake. By means of liquid chromatography consisting of gel filtration on Sephracryl S-200, S-300 and ion-exchange on DEAE Sepharose, leuc-B was purified to homogeneity. The pro...
Gespeichert in:
| Veröffentlicht in: | Archives of biochemistry and biophysics 2007-12, Vol.468 (2), p.193-204 |
|---|---|
| Hauptverfasser: | , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 204 |
|---|---|
| container_issue | 2 |
| container_start_page | 193 |
| container_title | Archives of biochemistry and biophysics |
| container_volume | 468 |
| creator | Sanchez, Eladio F. Gabriel, Lucilene M. Gontijo, Sileia Gremski, Luiza H. Veiga, Silvio S. Evangelista, Karla S. Eble, Johannes A. Richardson, Michael |
| description | Leucurolysin-B (leuc-B) is an hemorrhagic metalloproteinase found in the venom of
Bothrops leucurus (white-tailed-jararaca) snake. By means of liquid chromatography consisting of gel filtration on Sephracryl S-200, S-300 and ion-exchange on DEAE Sepharose, leuc-B was purified to homogeneity. The proteinase has an apparent molecular mass of 55
kDa as revealed by the reduced SDS–PAGE, and represents approximately 1.2% of the total protein in
B. leucurus venom. The partial amino acid sequence of leuc-B was determined by automated Edman sequencing of peptides derived from digests of the S-reduced and alkylated protein with trypsin. Leuc-B exhibits the characteristic motif of metalloproteinases, HEXXHXXGXXH and a methionine-containing turn of similar conformation (“Met-turn”), which forms a hydrophobic basis for the zinc ions and the three histidine residues involved as ligands. Leuc-B has been characterized as a P-III metalloproteinase and possesses a multidomain structure including a metalloproteinase, a disintegrin-like (ECD sequence instead of the typical RGD motif) and a cysteine-rich C-terminal domain. Leuc-B contains three potential sites of
N-glycosylation. The enzyme only cleaves the Ala
14–Leu
15 peptide bond of the oxidized insulin B-chain and preferentially hydrolyzes the Aα-chain of fibrinogen and the α-chain of fibrin. Its proteolytic activity was completely inhibited by metal chelating agents but not by other typical proteinase inhibitors. In addition, its enzymatic activity was stimulated by the divalent cations Ca
2+ and Mg
2+ but inhibited by Zn
2+ and Cu
2+. The catalytic activity of leuc-B on extracellular matrix proteins could readily lead to loss of capillary integrity resulting in hemorrhage occurring at those sites (MHD
=
30
ng in rabbit), with alterations in platelet function. In summary, here we report the isolation and the structure–function relationship of a P-III snake venom metalloproteinase. |
| doi_str_mv | 10.1016/j.abb.2007.10.002 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_68539508</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0003986107004973</els_id><sourcerecordid>68539508</sourcerecordid><originalsourceid>FETCH-LOGICAL-c417t-e3af6835e589c6f63c14c5fe3e00d7ae3e88000eafdc4205f2058888d0c661293</originalsourceid><addsrcrecordid>eNp9kE1rGzEQhkVoSJyPH9BL0aknrzNa7cpaeqpDmxoCLbQ5C1k7IjK7K1cfBufXR8aG3joghpl550XzEPKRwYIBEw_bhd5sFjXAstQLgPqCzBh0ogIumw9kBgC86qRg1-Qmxi0AY42or8g1W3aCC9nOSP6dQjYpBz1QPfXU5skk56dSmlcdtEkY3Js-tqi3VNNf1Xq9piMmPQx-F3xCN-mIczpgNjn44RDdVK3m1AY_0pVPr8Hv4nmaI93j5Mc7cmn1EPH-nG_Jy_dvfx5_VM8_n9aPX58r07BlqpBrKyRvsZWdEVZwwxrTWuQI0C91yVKWG1Hb3jQ1tLY8WaIHIwSrO35LPp98y0f_ZoxJjS4aHAY9oc9RFQS8a0EWITsJTfAxBrRqF9yow0ExUEfWaqsKa3VkfWwV1mXn09k8b0bs_22c4RbBl5MAy4l7h0FF43Ay2LuAJqneu__YvwNKG5DM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>68539508</pqid></control><display><type>article</type><title>Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Sanchez, Eladio F. ; Gabriel, Lucilene M. ; Gontijo, Sileia ; Gremski, Luiza H. ; Veiga, Silvio S. ; Evangelista, Karla S. ; Eble, Johannes A. ; Richardson, Michael</creator><creatorcontrib>Sanchez, Eladio F. ; Gabriel, Lucilene M. ; Gontijo, Sileia ; Gremski, Luiza H. ; Veiga, Silvio S. ; Evangelista, Karla S. ; Eble, Johannes A. ; Richardson, Michael</creatorcontrib><description>Leucurolysin-B (leuc-B) is an hemorrhagic metalloproteinase found in the venom of
Bothrops leucurus (white-tailed-jararaca) snake. By means of liquid chromatography consisting of gel filtration on Sephracryl S-200, S-300 and ion-exchange on DEAE Sepharose, leuc-B was purified to homogeneity. The proteinase has an apparent molecular mass of 55
kDa as revealed by the reduced SDS–PAGE, and represents approximately 1.2% of the total protein in
B. leucurus venom. The partial amino acid sequence of leuc-B was determined by automated Edman sequencing of peptides derived from digests of the S-reduced and alkylated protein with trypsin. Leuc-B exhibits the characteristic motif of metalloproteinases, HEXXHXXGXXH and a methionine-containing turn of similar conformation (“Met-turn”), which forms a hydrophobic basis for the zinc ions and the three histidine residues involved as ligands. Leuc-B has been characterized as a P-III metalloproteinase and possesses a multidomain structure including a metalloproteinase, a disintegrin-like (ECD sequence instead of the typical RGD motif) and a cysteine-rich C-terminal domain. Leuc-B contains three potential sites of
N-glycosylation. The enzyme only cleaves the Ala
14–Leu
15 peptide bond of the oxidized insulin B-chain and preferentially hydrolyzes the Aα-chain of fibrinogen and the α-chain of fibrin. Its proteolytic activity was completely inhibited by metal chelating agents but not by other typical proteinase inhibitors. In addition, its enzymatic activity was stimulated by the divalent cations Ca
2+ and Mg
2+ but inhibited by Zn
2+ and Cu
2+. The catalytic activity of leuc-B on extracellular matrix proteins could readily lead to loss of capillary integrity resulting in hemorrhage occurring at those sites (MHD
=
30
ng in rabbit), with alterations in platelet function. In summary, here we report the isolation and the structure–function relationship of a P-III snake venom metalloproteinase.</description><identifier>ISSN: 0003-9861</identifier><identifier>EISSN: 1096-0384</identifier><identifier>DOI: 10.1016/j.abb.2007.10.002</identifier><identifier>PMID: 17963685</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Bothrops - metabolism ; Bothrops leucurus ; Crotalid Venoms - chemistry ; Disintegrin-like protein ; Enzyme Activation ; Enzyme Stability ; Leucurolysin-B ; Metalloproteases - chemistry ; Metalloproteases - classification ; Metalloproteases - ultrastructure ; Metalloproteinase ; Molecular Sequence Data ; Snake venom</subject><ispartof>Archives of biochemistry and biophysics, 2007-12, Vol.468 (2), p.193-204</ispartof><rights>2007 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-e3af6835e589c6f63c14c5fe3e00d7ae3e88000eafdc4205f2058888d0c661293</citedby><cites>FETCH-LOGICAL-c417t-e3af6835e589c6f63c14c5fe3e00d7ae3e88000eafdc4205f2058888d0c661293</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.abb.2007.10.002$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,45974</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17963685$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sanchez, Eladio F.</creatorcontrib><creatorcontrib>Gabriel, Lucilene M.</creatorcontrib><creatorcontrib>Gontijo, Sileia</creatorcontrib><creatorcontrib>Gremski, Luiza H.</creatorcontrib><creatorcontrib>Veiga, Silvio S.</creatorcontrib><creatorcontrib>Evangelista, Karla S.</creatorcontrib><creatorcontrib>Eble, Johannes A.</creatorcontrib><creatorcontrib>Richardson, Michael</creatorcontrib><title>Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom</title><title>Archives of biochemistry and biophysics</title><addtitle>Arch Biochem Biophys</addtitle><description>Leucurolysin-B (leuc-B) is an hemorrhagic metalloproteinase found in the venom of
Bothrops leucurus (white-tailed-jararaca) snake. By means of liquid chromatography consisting of gel filtration on Sephracryl S-200, S-300 and ion-exchange on DEAE Sepharose, leuc-B was purified to homogeneity. The proteinase has an apparent molecular mass of 55
kDa as revealed by the reduced SDS–PAGE, and represents approximately 1.2% of the total protein in
B. leucurus venom. The partial amino acid sequence of leuc-B was determined by automated Edman sequencing of peptides derived from digests of the S-reduced and alkylated protein with trypsin. Leuc-B exhibits the characteristic motif of metalloproteinases, HEXXHXXGXXH and a methionine-containing turn of similar conformation (“Met-turn”), which forms a hydrophobic basis for the zinc ions and the three histidine residues involved as ligands. Leuc-B has been characterized as a P-III metalloproteinase and possesses a multidomain structure including a metalloproteinase, a disintegrin-like (ECD sequence instead of the typical RGD motif) and a cysteine-rich C-terminal domain. Leuc-B contains three potential sites of
N-glycosylation. The enzyme only cleaves the Ala
14–Leu
15 peptide bond of the oxidized insulin B-chain and preferentially hydrolyzes the Aα-chain of fibrinogen and the α-chain of fibrin. Its proteolytic activity was completely inhibited by metal chelating agents but not by other typical proteinase inhibitors. In addition, its enzymatic activity was stimulated by the divalent cations Ca
2+ and Mg
2+ but inhibited by Zn
2+ and Cu
2+. The catalytic activity of leuc-B on extracellular matrix proteins could readily lead to loss of capillary integrity resulting in hemorrhage occurring at those sites (MHD
=
30
ng in rabbit), with alterations in platelet function. In summary, here we report the isolation and the structure–function relationship of a P-III snake venom metalloproteinase.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Bothrops - metabolism</subject><subject>Bothrops leucurus</subject><subject>Crotalid Venoms - chemistry</subject><subject>Disintegrin-like protein</subject><subject>Enzyme Activation</subject><subject>Enzyme Stability</subject><subject>Leucurolysin-B</subject><subject>Metalloproteases - chemistry</subject><subject>Metalloproteases - classification</subject><subject>Metalloproteases - ultrastructure</subject><subject>Metalloproteinase</subject><subject>Molecular Sequence Data</subject><subject>Snake venom</subject><issn>0003-9861</issn><issn>1096-0384</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1rGzEQhkVoSJyPH9BL0aknrzNa7cpaeqpDmxoCLbQ5C1k7IjK7K1cfBufXR8aG3joghpl550XzEPKRwYIBEw_bhd5sFjXAstQLgPqCzBh0ogIumw9kBgC86qRg1-Qmxi0AY42or8g1W3aCC9nOSP6dQjYpBz1QPfXU5skk56dSmlcdtEkY3Js-tqi3VNNf1Xq9piMmPQx-F3xCN-mIczpgNjn44RDdVK3m1AY_0pVPr8Hv4nmaI93j5Mc7cmn1EPH-nG_Jy_dvfx5_VM8_n9aPX58r07BlqpBrKyRvsZWdEVZwwxrTWuQI0C91yVKWG1Hb3jQ1tLY8WaIHIwSrO35LPp98y0f_ZoxJjS4aHAY9oc9RFQS8a0EWITsJTfAxBrRqF9yow0ExUEfWaqsKa3VkfWwV1mXn09k8b0bs_22c4RbBl5MAy4l7h0FF43Ay2LuAJqneu__YvwNKG5DM</recordid><startdate>20071215</startdate><enddate>20071215</enddate><creator>Sanchez, Eladio F.</creator><creator>Gabriel, Lucilene M.</creator><creator>Gontijo, Sileia</creator><creator>Gremski, Luiza H.</creator><creator>Veiga, Silvio S.</creator><creator>Evangelista, Karla S.</creator><creator>Eble, Johannes A.</creator><creator>Richardson, Michael</creator><general>Elsevier Inc</general><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>20071215</creationdate><title>Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom</title><author>Sanchez, Eladio F. ; Gabriel, Lucilene M. ; Gontijo, Sileia ; Gremski, Luiza H. ; Veiga, Silvio S. ; Evangelista, Karla S. ; Eble, Johannes A. ; Richardson, Michael</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-e3af6835e589c6f63c14c5fe3e00d7ae3e88000eafdc4205f2058888d0c661293</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Bothrops - metabolism</topic><topic>Bothrops leucurus</topic><topic>Crotalid Venoms - chemistry</topic><topic>Disintegrin-like protein</topic><topic>Enzyme Activation</topic><topic>Enzyme Stability</topic><topic>Leucurolysin-B</topic><topic>Metalloproteases - chemistry</topic><topic>Metalloproteases - classification</topic><topic>Metalloproteases - ultrastructure</topic><topic>Metalloproteinase</topic><topic>Molecular Sequence Data</topic><topic>Snake venom</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sanchez, Eladio F.</creatorcontrib><creatorcontrib>Gabriel, Lucilene M.</creatorcontrib><creatorcontrib>Gontijo, Sileia</creatorcontrib><creatorcontrib>Gremski, Luiza H.</creatorcontrib><creatorcontrib>Veiga, Silvio S.</creatorcontrib><creatorcontrib>Evangelista, Karla S.</creatorcontrib><creatorcontrib>Eble, Johannes A.</creatorcontrib><creatorcontrib>Richardson, Michael</creatorcontrib><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>Sanchez, Eladio F.</au><au>Gabriel, Lucilene M.</au><au>Gontijo, Sileia</au><au>Gremski, Luiza H.</au><au>Veiga, Silvio S.</au><au>Evangelista, Karla S.</au><au>Eble, Johannes A.</au><au>Richardson, Michael</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom</atitle><jtitle>Archives of biochemistry and biophysics</jtitle><addtitle>Arch Biochem Biophys</addtitle><date>2007-12-15</date><risdate>2007</risdate><volume>468</volume><issue>2</issue><spage>193</spage><epage>204</epage><pages>193-204</pages><issn>0003-9861</issn><eissn>1096-0384</eissn><abstract>Leucurolysin-B (leuc-B) is an hemorrhagic metalloproteinase found in the venom of
Bothrops leucurus (white-tailed-jararaca) snake. By means of liquid chromatography consisting of gel filtration on Sephracryl S-200, S-300 and ion-exchange on DEAE Sepharose, leuc-B was purified to homogeneity. The proteinase has an apparent molecular mass of 55
kDa as revealed by the reduced SDS–PAGE, and represents approximately 1.2% of the total protein in
B. leucurus venom. The partial amino acid sequence of leuc-B was determined by automated Edman sequencing of peptides derived from digests of the S-reduced and alkylated protein with trypsin. Leuc-B exhibits the characteristic motif of metalloproteinases, HEXXHXXGXXH and a methionine-containing turn of similar conformation (“Met-turn”), which forms a hydrophobic basis for the zinc ions and the three histidine residues involved as ligands. Leuc-B has been characterized as a P-III metalloproteinase and possesses a multidomain structure including a metalloproteinase, a disintegrin-like (ECD sequence instead of the typical RGD motif) and a cysteine-rich C-terminal domain. Leuc-B contains three potential sites of
N-glycosylation. The enzyme only cleaves the Ala
14–Leu
15 peptide bond of the oxidized insulin B-chain and preferentially hydrolyzes the Aα-chain of fibrinogen and the α-chain of fibrin. Its proteolytic activity was completely inhibited by metal chelating agents but not by other typical proteinase inhibitors. In addition, its enzymatic activity was stimulated by the divalent cations Ca
2+ and Mg
2+ but inhibited by Zn
2+ and Cu
2+. The catalytic activity of leuc-B on extracellular matrix proteins could readily lead to loss of capillary integrity resulting in hemorrhage occurring at those sites (MHD
=
30
ng in rabbit), with alterations in platelet function. In summary, here we report the isolation and the structure–function relationship of a P-III snake venom metalloproteinase.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>17963685</pmid><doi>10.1016/j.abb.2007.10.002</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-9861 |
| ispartof | Archives of biochemistry and biophysics, 2007-12, Vol.468 (2), p.193-204 |
| issn | 0003-9861 1096-0384 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_68539508 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Amino Acid Sequence Animals Bothrops - metabolism Bothrops leucurus Crotalid Venoms - chemistry Disintegrin-like protein Enzyme Activation Enzyme Stability Leucurolysin-B Metalloproteases - chemistry Metalloproteases - classification Metalloproteases - ultrastructure Metalloproteinase Molecular Sequence Data Snake venom |
| title | Structural and functional characterization of a P-III metalloproteinase, leucurolysin-B, from Bothrops leucurus venom |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-21T12%3A49%3A59IST&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=Structural%20and%20functional%20characterization%20of%20a%20P-III%20metalloproteinase,%20leucurolysin-B,%20from%20Bothrops%20leucurus%20venom&rft.jtitle=Archives%20of%20biochemistry%20and%20biophysics&rft.au=Sanchez,%20Eladio%20F.&rft.date=2007-12-15&rft.volume=468&rft.issue=2&rft.spage=193&rft.epage=204&rft.pages=193-204&rft.issn=0003-9861&rft.eissn=1096-0384&rft_id=info:doi/10.1016/j.abb.2007.10.002&rft_dat=%3Cproquest_cross%3E68539508%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=68539508&rft_id=info:pmid/17963685&rft_els_id=S0003986107004973&rfr_iscdi=true |