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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.
MedLine Citation:
PMID:  23547002     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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.
Authors:
Kouhei Nagai; Koichi Morimoto; Haruka Ikegami; Hajime Kimura; Norishige Yotsukura
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-4-3
Journal Detail:
Title:  Marine biotechnology (New York, N.Y.)     Volume:  -     ISSN:  1436-2236     ISO Abbreviation:  Mar. Biotechnol.     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-4-2     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100892712     Medline TA:  Mar Biotechnol (NY)     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Genetic Engineering, Faculity of Biology-Oriented Science and Technology, Kinki University, Kinokawa, Wakayama, 649-6493, Japan.
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