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| 2. |
- Atteia, A., et al.
(author)
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Structure, organization and expression of the genes encoding mitochondrial cytochrome c1 and the Rieske iron-sulfur protein in Chlamydomonas reinhardtii
- 2003
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In: Molecular Genetics and Genomics. - : Springer Verlag. - 1617-4615 .- 1617-4623. ; 268:5, s. 637-644
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Journal article (peer-reviewed)abstract
- The sequence and organization of the Chlamydomonas reinhardtii genes encoding cytochrome c 1 ( Cyc1) and the Rieske-type iron-sulfur protein ( Isp), two key nucleus-encoded subunits of the mitochondrial cytochrome bc 1 complex, are presented. Southern hybridization analysis indicates that both Cyc1 and Isp are present as single-copy genes in C. reinhardtii. The Cyc1 gene spans 6404 bp and contains six introns, ranging from 178 to 1134 bp in size. The Isp gene spans 1238 bp and contains four smaller introns, ranging in length from 83 to 167 bp. In both genes, the intron/exon junctions follow the GT/AG rule. Internal conserved sequences were identified in only some of the introns in the Cyc1 gene. The levels of expression of Isp and Cyc1 genes are comparable in wild-type C. reinhardtii cells and in a mutant strain carrying a deletion in the mitochondrial gene for cytochrome b (dum-1). Nevertheless, no accumulation of the nucleus-encoded cytochrome c 1 or of core proteins I and II was observed in the membranes of the respiratory mutant. These data show that, in the green alga C. reinhardtii, the subunits of the cytochrome bc1 complex fail to assemble properly in the absence of cytochrome b.
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| 3. |
- Bohlin, Jan
(author)
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Enzymes and electron transport in microbial chlorate respiration
- 2008
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Doctoral thesis (other academic/artistic)abstract
- Microbial chlorate respiration plays an important role in the turnover of oxochlorates in nature and industrial waste management. This thesis deals with the characterization of the molecular components of chlorate respiration in Ideonella dechloratans. Chlorate respiration utilizes two soluble periplasmic enzymes, chlorate reductase and chlorite dismutase, to convert chlorate to chloride and oxygen. The genes encoding the enzymes participating in the chlorate degradation have been sequenced, and are found in close proximity, forming a gene cluster for chlorate metabolism. This work also includes the successful recombinant expression of three genes from Ideonella dechloratans. Two of the gene products, chlorite dismutase and the C subunit of chlorate reductase, participate in the chlorate respiration. The third gene, which is found close to the gene cluster for chlorate metabolism, encodes a soluble c-type cytochrome. The localization of the gene suggests the corresponding protein as a candidate for a role as electron donor to chlorate reductase. Also, the role of soluble periplasmic c cytochromes of Ideonella dechloratans in chlorate respiration was studied. At least one of the soluble c cytochromes was found capable of serving as electron donor for chlorate reduction. This c cytochrome, and several others, can also donate electrons to a terminal oxidase for subsequent reduction of oxygen, as required for the branched electron flow during chlorate respiration.
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| 4. |
- Cardol, Pierre, et al.
(author)
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Oxidative Phosphorylation : Building blocks and related components
- 2009
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In: The Chlamydomonas Sourcebook. - Oxford : Academic Press. - 0123708753 ; , s. 469-502
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Book chapter (other academic/artistic)abstract
- This chapter summarizes the knowledge of the oxidative phosphorylation (OXPHOS) constituents of Chlamydomonas and of the components involved in their biogenesis and addresses alternate dehydrogenases and oxidases which are particular to photosynthetic organisms, and several other mitochondrial components related to OXPHOS. Reference to the components of Polytomella sp., a colorless alga closely related to Chlamydomonas is clearly made. The main complexes involved in electron transport seem to share a similar number of subunits, and many of the algal polypeptides have plant homologues. Some differences are apparent, such as the presence of a fragmented COX2 subunit, which seems to be unique to chlorophyte algae. OXPHOS is defined as an electron transfer chain driven by substrate oxidation that is coupled to the synthesis of ATP through an electrochemical transmembrane gradient. The characterization of Arabidopsis mitochondrial components through proteomic approaches has advanced significantly. As a unicellular organism, Chlamydomonas offers the unique opportunity to study organelle-organelle interactions, particularly between mitochondria and chloroplasts. It has become evident that crosstalk between these organelles takes place, mainly through intracellular metabolite pools. © 2009 Elsevier Inc. All rights reserved.
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| 5. |
- Eriksson, Gunnar, et al.
(author)
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Exploiting Syntax when Detecting Protein Names in Text
- 2002. - 1
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In: Proceedings of FMI Workshop on Natural Language Processing in Biomedical Applications.
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Conference paper (peer-reviewed)abstract
- This paper presents work on a method to detect names of proteins in running text. Our system - Yapex - uses a combination of lexical and syntactic knowledge, heuristic filters and a local dynamic dictionary. The syntactic information given by a general-purpose off-the-shelf parser supports the correct identification of the boundaries of protein names, and the local dynamic dictionary finds protein names in positions incompletely analysed by the parser. We present the different steps involved in our approach to protein tagging, and show how combinations of them influence recall and precision. We evaluate the system on a corpus of MEDLINE abstracts and compare it with the KeX system (Fukuda et al., 1998) along four different notions of correctness.
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| 6. |
- Eriksson, Martin, 1970, et al.
(author)
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Community-Level Analysis of psbA Gene Sequences and Irgarol Tolerance in Marine Periphyton
- 2009
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In: Applied and Environmental Microbiology. - Washington, D.C. : American Society for Microbiology. - 0099-2240 .- 1098-5336. ; 75:4, s. 897-906
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Journal article (peer-reviewed)abstract
- This study analyzes psbA gene sequences, predicted D1 protein sequences, species relative abundance, and pollution-induced community tolerance in marine periphyton communities exposed to the antifouling compound Irgarol 1051. The mechanism of action of Irgarol is the inhibition of photosynthetic electron transport at photosystem II by binding to the D1 protein. The metagenome of the communities was used to produce clone libraries containing fragments of the psbA gene encoding the D1 protein. Community tolerance was quantified with a short-term test for the inhibition of photosynthesis. The communities were established in a continuous flow of natural seawater through microcosms with or without added Irgarol. The selection pressure from Irgarol resulted in an altered species composition and an inducted community tolerance to Irgarol. Moreover, there was a very high diversity in the psbA gene sequences in the periphyton, and the composition of psbA and D1 fragments within the communities was dramatically altered by increased Irgarol exposure. Even though tolerance to this type of compound in land plants often depends on a single amino acid substitution (Ser(264)-> Gly) in the D1 protein, this was not the case for marine periphyton species. Instead, the tolerance mechanism likely involves increased degradation of D1. When we compared sequences from low and high Irgarol exposure, differences in nonconserved amino acids were found only in the so-called PEST region of D1, which is involved in regulating its degradation. Our results suggest that environmental contamination with Irgarol has led to selection for high-turnover D1 proteins in marine periphyton communities at the west coast of Sweden.
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| 7. |
- Farkas, Daniel, et al.
(author)
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Cloning, expression and purification of the luminal domain of spinach photosystem 1 subunit PsaF functional in binding to plastocyanin and with a disulfide bridge required for folding
- 2011
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In: Protein Expression and Purification. - San Diego : Academic Press. - 1046-5928 .- 1096-0279. ; 78:2, s. 156-166
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Journal article (peer-reviewed)abstract
- The photosystem 1 subunit PsaF is involved in the docking of the electron-donor proteins plastocyanin and cytochrome c6 in eukaryotic photosynthetic organisms. Here we report the expression, purification and basic characterization of the luminal domain of spinach PsaF, encompassing amino-acid residues 1-79. The recombinant protein was expressed in Escherichia coli BL21 (DE3) using a pET32 Xa/LIC thioredoxin fusion system. The thioredoxin fusion protein contained a His6 tag and was removed and separated from PsaF through proteolytic digestion by factor Xa followed by immobilized metal affinity chromatography. Further purification with size-exclusion chromatography resulted in a final yield of approximately 6 mg PsaF from one liter growth medium. The correct identity after the factor Xa treatment of PsaF was verified by FT-ICR mass spectrometry which also showed that the purified protein contains an intact disulfide bridge between Cys residues 6 and 38. Secondary structure and folding was further explored using far-UV CD spectroscopy indicating a α-helical content in agreement with the 3.3 Å-resolution crystal structure of photosystem I Ref. [5] and a helix-coil transition temperature of 29 °C. Thermofluorescence studies showed that the disulfide bridge is necessary to keep the overall fold of the protein and that hydrophobic regions become exposed at 50-65 °C depending on the ionic strength. The described expression and purification procedure can be used for isotopic labeling of the protein and 15N-HSQC NMR studies indicated a slow or intermediate exchange between different conformations of the prepared protein and that it belongs to the molten-globule structural family. Finally, by using a carboxyl- and amine-reactive zero-length crosslinker, we have shown that the recombinant protein binds to plastocyanin by a specific, native-like, electrostatic interaction, hence, confirming its functionality.
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| 8. |
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| 9. |
- Figueroa-Martinez, Francisco, et al.
(author)
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Reconstructing the mitochondrial protein import machinery of Chlamydomonas reinhardtii
- 2008
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In: Genetics. - Austin, Tex. : The Genetics Society. - 0016-6731 .- 1943-2631. ; 179:1, s. 149-155
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Journal article (peer-reviewed)abstract
- In Chlamydomonas reinhardtii several nucleus-encoded proteins that participate in the mitochondrial oxidative phosphorylation are targeted to the organelle by unusually long mitochondrial targeting sequences. Here, we explored the components of the mitochondrial import machinery of the green alga. We mined the algal genome, searching for yeast and plant homologs, and reconstructed the mitochondrial import machinery. All the main translocation components were identified in Chlamydomonas as well as in Arabidopsis thaliana and in the recently sequenced moss Physcomitrella patens. Some of these components appear to be duplicated, as is the case of Tim22. In contrast, several yeast components that have relatively large hydrophilic regions exposed to the cytosol or to the intermembrane space seem to be absent in land plants and green algae. If present at all, these components of plants and algae may differ significantly from their yeast counterparts. We propose that long mitochondrial targeting sequences in some Chlamydomonas mitochondrial protein precursors are involved in preventing the aggregation of the hydrophobic proteins they carry.
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| 10. |
- Franzén, Kristofer, et al.
(author)
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Protein names and how to find them
- 2002. - 1
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In: International Journal of Medical Informatics. - : Elsevier. - 1386-5056 .- 1872-8243. ; 67, s. 49-61
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Journal article (peer-reviewed)abstract
- A prerequisite for all higher level information extraction tasks is the identification of unknown names in text. Today, when large corpora can consist of billions of words, it is of utmost importance to develop accurate techniques for the automatic detection, extraction and categorization of named entities in these corpora. Although named entity recognition might be regarded a solved problem in some domains, it still poses a significant challenge in others. In this work we focus on one of the more difficult tasks, the identification of protein names in text. This task presents several interesting difficulties because of the named entities' variant structural characteristics, their sometimes unclear status as names, the lack of common standards and fixed nomenclatures, and the specifics of the texts in the molecular biology domain in which they appear. We describe how we approached these and other difficulties in the implementation of Yapex, a system for the automatic identification of protein names in text. We also evaluate Yapex under four different notions of correctness and compare its performance to that of another publicly available system for protein name recognition.
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