Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495.
E495 is the most abundant protease secreted by the Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. As a thermolysin family metalloprotease, E495 was found to have multiple active forms in the culture of strain SM495. E495-M (containing only the catalytic domain) and E495-M-C1 (containing the c...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22523598/pdf/?tool=EBI |
| _version_ | 1818337721689571328 |
|---|---|
| author | Hai-Lun He Jun Guo Xiu-Lan Chen Bin-Bin Xie Xi-Ying Zhang Yong Yu Bo Chen Bai-Cheng Zhou Yu-Zhong Zhang |
| author_facet | Hai-Lun He Jun Guo Xiu-Lan Chen Bin-Bin Xie Xi-Ying Zhang Yong Yu Bo Chen Bai-Cheng Zhou Yu-Zhong Zhang |
| author_sort | Hai-Lun He |
| collection | DOAJ |
| description | E495 is the most abundant protease secreted by the Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. As a thermolysin family metalloprotease, E495 was found to have multiple active forms in the culture of strain SM495. E495-M (containing only the catalytic domain) and E495-M-C1 (containing the catalytic domain and one PPC domain) were two stable mature forms, and E495-M-C1-C2 (containing the catalytic domain and two PPC domains) might be an intermediate. Compared to E495-M, E495-M-C1 had similar affinity and catalytic efficiency to oligopeptides, but higher affinity and catalytic efficiency to proteins. The PPC domains from E495 were expressed as GST-fused proteins. Both of the recombinant PPC domains were shown to have binding ability to proteins C-phycocyanin and casein, and domain PPC1 had higher affinity to C-phycocyanin than domain PPC2. These results indicated that the domain PPC1 in E495-M-C1 could be helpful in binding protein substrate, and therefore, improving the catalytic efficiency. Site-directed mutagenesis on the PPC domains showed that the conserved polar and aromatic residues, D26, D28, Y30, Y/W65, in the PPC domains played key roles in protein binding. Our study may shed light on the mechanism of organic nitrogen degradation in the Arctic sea ice. |
| first_indexed | 2024-12-13T14:59:43Z |
| format | Article |
| id | doaj.art-5f7e7309ea5247c682967d2f1d4fc420 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T14:59:43Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-5f7e7309ea5247c682967d2f1d4fc4202022-12-21T23:41:09ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0174e3544210.1371/journal.pone.0035442Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495.Hai-Lun HeJun GuoXiu-Lan ChenBin-Bin XieXi-Ying ZhangYong YuBo ChenBai-Cheng ZhouYu-Zhong ZhangE495 is the most abundant protease secreted by the Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. As a thermolysin family metalloprotease, E495 was found to have multiple active forms in the culture of strain SM495. E495-M (containing only the catalytic domain) and E495-M-C1 (containing the catalytic domain and one PPC domain) were two stable mature forms, and E495-M-C1-C2 (containing the catalytic domain and two PPC domains) might be an intermediate. Compared to E495-M, E495-M-C1 had similar affinity and catalytic efficiency to oligopeptides, but higher affinity and catalytic efficiency to proteins. The PPC domains from E495 were expressed as GST-fused proteins. Both of the recombinant PPC domains were shown to have binding ability to proteins C-phycocyanin and casein, and domain PPC1 had higher affinity to C-phycocyanin than domain PPC2. These results indicated that the domain PPC1 in E495-M-C1 could be helpful in binding protein substrate, and therefore, improving the catalytic efficiency. Site-directed mutagenesis on the PPC domains showed that the conserved polar and aromatic residues, D26, D28, Y30, Y/W65, in the PPC domains played key roles in protein binding. Our study may shed light on the mechanism of organic nitrogen degradation in the Arctic sea ice.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22523598/pdf/?tool=EBI |
| spellingShingle | Hai-Lun He Jun Guo Xiu-Lan Chen Bin-Bin Xie Xi-Ying Zhang Yong Yu Bo Chen Bai-Cheng Zhou Yu-Zhong Zhang Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. PLoS ONE |
| title | Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. |
| title_full | Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. |
| title_fullStr | Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. |
| title_full_unstemmed | Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. |
| title_short | Structural and functional characterization of mature forms of metalloprotease E495 from Arctic sea-ice bacterium Pseudoalteromonas sp. SM495. |
| title_sort | structural and functional characterization of mature forms of metalloprotease e495 from arctic sea ice bacterium pseudoalteromonas sp sm495 |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22523598/pdf/?tool=EBI |
| work_keys_str_mv | AT hailunhe structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT junguo structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT xiulanchen structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT binbinxie structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT xiyingzhang structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT yongyu structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT bochen structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT baichengzhou structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 AT yuzhongzhang structuralandfunctionalcharacterizationofmatureformsofmetalloproteasee495fromarcticseaicebacteriumpseudoalteromonasspsm495 |