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Investigation of Proteomic Profiles of Lamina of Ecklonia kurome (Laminariales): Homology-Based Cross-Species Protein Identification and Analysis of the Post-translational Processing of Vanadium-Dependent Bromoperoxidases Using MALDI-TOF/TOF
| Title: | Investigation of Proteomic Profiles of Lamina of Ecklonia kurome (Laminariales): Homology-Based Cross-Species Protein Identification and Analysis of the Post-translational Processing of Vanadium-Dependent Bromoperoxidases Using MALDI-TOF/TOF |
| Authors: | Nagai, Kouhei Browse this author | | Morimoto, Koichi Browse this author →KAKEN DB | | Ikegami, Haruka Browse this author | | Kimura, Hajime Browse this author | | Yotsukura, Norishige Browse this author →KAKEN DB |
| Keywords: | 2-DE | | Kelp | | MALDI-TOF/TOF | | Protein identification | | Haloperoxidase |
| Issue Date: | Aug-2013 |
| Publisher: | Springer |
| Journal Title: | Marine Biotechnology |
| Volume: | 15 |
| Issue: | 4 |
| Start Page: | 487 |
| End Page: | 498 |
| Publisher DOI: | 10.1007/s10126-013-9498-z |
| Abstract: | Proteomic profiles of the lamina of Ecklonia kurome Okamura, one of the Japanese dominant laminarialean kelps, were investigated by two-dimensional electrophoresis (2-DE) and MALDI-TOF/TOF. Due to the absence of E. kurome DNA or protein databases, homology-based cross-species protein identification was performed using a combination of three database-searching algorithms, Mascot peptide mass fingerprinting, Mascot MS/MS ion search, and mass spectrometry-based BLAST. Proteins were extracted from the lamina by an ethanol/phenol method and subjected to 2-DE (pI 4-7, 10 % polyacrylamide gel). More than 700 spots were detected in the 2-DE gel with CBB, and 93 spots (24 proteins) were successfully identified by MALDI-TOF/TOF and the cross-species database searching. The identified proteins mainly consisted of cytoplasmic carbohydrate metabolic enzymes, chloroplast proteins involved in photosynthesis, and haloperoxidases. Interestingly, vanadium-dependent bromoperoxidases (vBPO), which is thought to be involved in halogen uptake, synthesis of halogenated products, and detoxification of reactive oxygen species, were separated into at least 23 different spots. By comparing mass spectra, amino acid sequences predicted from tandem mass spectra and haloperoxidase activities of the vBPOs, we found that (1) at least two types of vBPOs were expressed in the lamina of E. kurome and (2) two pro-vBPOs might be activated by specific cleavage at N- and C-terminal regions. |
| Rights: | The final publication is available at link.springer.com. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/56644 |
| Appears in Collections: | 北方生物圏フィールド科学センター (Field Science Center for Northern Biosphere) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 四ツ倉 典滋
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