Assembly of Rieske iron-sulphur protein into the cytochrome bf complex in thylakoid membranes of isolated pea chloroplasts
The assembly of the Rieske iron–sulphur protein into the cytochrome bf complex was examined following import of 35S‐labeled precursor protein by isolated pea chloroplasts. Rieske protein assembled into the cytochrome bf complex was resolved from unassembled Rieske protein and from other membrane com...
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| Veröffentlicht in: | European journal of biochemistry 2000, Vol.267 (2), p.352-360 |
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| creator | Kapazoglou, A Mould, R.M Gray, J.C |
| description | The assembly of the Rieske iron–sulphur protein into the cytochrome bf complex was examined following import of 35S‐labeled precursor protein by isolated pea chloroplasts. Rieske protein assembled into the cytochrome bf complex was resolved from unassembled Rieske protein and from other membrane complexes by nondenaturing gel electrophoresis of dodecyl maltoside‐solubilized thylakoid membranes. Four mutant forms of the Rieske protein were able to assemble into the cytochrome bf complex in isolated chloroplasts. These were a triple substitution mutant, C107S/H109R/C112S, replacing conserved residues involved in the ligation of the [2Fe‐2S] centre; the mutant Δ45–52 which removed a glycine‐rich region predicted to form a flexible hinge between the hydrophobic membrane‐associated region and the hydrophilic lumenal domain; and mutants Δ168–173 and Δ177–179 which removed two C‐terminal regions, which are highly conserved in chloroplast and cyanobacterial Rieske proteins. This indicates that the [2Fe–2S] cluster, the glycine‐rich region and the C‐terminal region are not essential for stable assembly of the Rieske protein into the cytochrome bf complex in isolated chloroplasts. |
| doi_str_mv | 10.1046/j.1432-1327.2000.01006.x |
| format | Article |
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Rieske protein assembled into the cytochrome bf complex was resolved from unassembled Rieske protein and from other membrane complexes by nondenaturing gel electrophoresis of dodecyl maltoside‐solubilized thylakoid membranes. Four mutant forms of the Rieske protein were able to assemble into the cytochrome bf complex in isolated chloroplasts. These were a triple substitution mutant, C107S/H109R/C112S, replacing conserved residues involved in the ligation of the [2Fe‐2S] centre; the mutant Δ45–52 which removed a glycine‐rich region predicted to form a flexible hinge between the hydrophobic membrane‐associated region and the hydrophilic lumenal domain; and mutants Δ168–173 and Δ177–179 which removed two C‐terminal regions, which are highly conserved in chloroplast and cyanobacterial Rieske proteins. 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Rieske protein assembled into the cytochrome bf complex was resolved from unassembled Rieske protein and from other membrane complexes by nondenaturing gel electrophoresis of dodecyl maltoside‐solubilized thylakoid membranes. Four mutant forms of the Rieske protein were able to assemble into the cytochrome bf complex in isolated chloroplasts. These were a triple substitution mutant, C107S/H109R/C112S, replacing conserved residues involved in the ligation of the [2Fe‐2S] centre; the mutant Δ45–52 which removed a glycine‐rich region predicted to form a flexible hinge between the hydrophobic membrane‐associated region and the hydrophilic lumenal domain; and mutants Δ168–173 and Δ177–179 which removed two C‐terminal regions, which are highly conserved in chloroplast and cyanobacterial Rieske proteins. This indicates that the [2Fe–2S] cluster, the glycine‐rich region and the C‐terminal region are not essential for stable assembly of the Rieske protein into the cytochrome bf complex in isolated chloroplasts.</description><subject>assembly</subject><subject>Biological Transport</subject><subject>Cell Membrane - chemistry</subject><subject>Cell Membrane - metabolism</subject><subject>chloroplast</subject><subject>Chloroplasts - metabolism</subject><subject>Cytochrome b Group - biosynthesis</subject><subject>Cytochrome b Group - metabolism</subject><subject>Cytochrome b6f Complex</subject><subject>cytochrome bf complex</subject><subject>Electron Transport Complex III</subject><subject>Electrophoresis - methods</subject><subject>Iron-Sulfur Proteins - genetics</subject><subject>Iron-Sulfur Proteins - metabolism</subject><subject>Mutation</subject><subject>Pisum sativum</subject><subject>Pisum sativum - metabolism</subject><subject>Rieske FeS protein</subject><subject>thylakoid membrane</subject><subject>thylakoids</subject><subject>Thylakoids - chemistry</subject><subject>Thylakoids - metabolism</subject><subject>Time Factors</subject><issn>0014-2956</issn><issn>1432-1033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkMFu1DAURS0EotPCL4BX7BKeYztONkilagtSJSRa1paTvDCeOuNgJ-qEr8chXbBkZUvv3uPnQwhlkDMQ5cdDzgQvMsYLlRcAkAMDKPPTC7LbBsD5S7IDYCIralmekfMYDylY1qV6Tc4YlKkKckd-X8aIQ-MW6nv63WJ8RGqDP2ZxduN-DnQMfkJ7pPY4eTrtkbbL5Nt98APSpqetH0aHpzROw8WZR287OiRiMEeMK9RG78yEHR3R0HbvfPCjM3GKb8ir3riIb5_PC_Jwc_1w9SW7-3b79eryLmuFgDJTlVR1V7aFbAWXXc8rYNh1CrFuKgApCylVJSSXNTeVwa7ulcFasaZn0El-QT5s2PSTXzPGSQ82tuhcWtDPUSuoZCVFmYLVFmyDjzFgr8dgBxMWzUCv2vVBr3b1ql2v2vVf7fqUqu-e35ibAbt_ipvnFPi0BZ6sw-W_wfrm-vP9ek2A9xugN16bn8FG_eO-AMahqEUllOR_ACEsnAM</recordid><startdate>2000</startdate><enddate>2000</enddate><creator>Kapazoglou, A</creator><creator>Mould, R.M</creator><creator>Gray, J.C</creator><general>Blackwell Science Ltd</general><scope>FBQ</scope><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>2000</creationdate><title>Assembly of Rieske iron-sulphur protein into the cytochrome bf complex in thylakoid membranes of isolated pea chloroplasts</title><author>Kapazoglou, A ; Mould, R.M ; Gray, J.C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4406-78579d6c25c435df3801edd7ee9b8005525578453593a8aed9f7ae971bf10d53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>assembly</topic><topic>Biological Transport</topic><topic>Cell Membrane - chemistry</topic><topic>Cell Membrane - metabolism</topic><topic>chloroplast</topic><topic>Chloroplasts - metabolism</topic><topic>Cytochrome b Group - biosynthesis</topic><topic>Cytochrome b Group - metabolism</topic><topic>Cytochrome b6f Complex</topic><topic>cytochrome bf complex</topic><topic>Electron Transport Complex III</topic><topic>Electrophoresis - methods</topic><topic>Iron-Sulfur Proteins - genetics</topic><topic>Iron-Sulfur Proteins - metabolism</topic><topic>Mutation</topic><topic>Pisum sativum</topic><topic>Pisum sativum - metabolism</topic><topic>Rieske FeS protein</topic><topic>thylakoid membrane</topic><topic>thylakoids</topic><topic>Thylakoids - chemistry</topic><topic>Thylakoids - metabolism</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kapazoglou, A</creatorcontrib><creatorcontrib>Mould, R.M</creatorcontrib><creatorcontrib>Gray, J.C</creatorcontrib><collection>AGRIS</collection><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>European journal of biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kapazoglou, A</au><au>Mould, R.M</au><au>Gray, J.C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Assembly of Rieske iron-sulphur protein into the cytochrome bf complex in thylakoid membranes of isolated pea chloroplasts</atitle><jtitle>European journal of biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>2000</date><risdate>2000</risdate><volume>267</volume><issue>2</issue><spage>352</spage><epage>360</epage><pages>352-360</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><abstract>The assembly of the Rieske iron–sulphur protein into the cytochrome bf complex was examined following import of 35S‐labeled precursor protein by isolated pea chloroplasts. Rieske protein assembled into the cytochrome bf complex was resolved from unassembled Rieske protein and from other membrane complexes by nondenaturing gel electrophoresis of dodecyl maltoside‐solubilized thylakoid membranes. Four mutant forms of the Rieske protein were able to assemble into the cytochrome bf complex in isolated chloroplasts. These were a triple substitution mutant, C107S/H109R/C112S, replacing conserved residues involved in the ligation of the [2Fe‐2S] centre; the mutant Δ45–52 which removed a glycine‐rich region predicted to form a flexible hinge between the hydrophobic membrane‐associated region and the hydrophilic lumenal domain; and mutants Δ168–173 and Δ177–179 which removed two C‐terminal regions, which are highly conserved in chloroplast and cyanobacterial Rieske proteins. This indicates that the [2Fe–2S] cluster, the glycine‐rich region and the C‐terminal region are not essential for stable assembly of the Rieske protein into the cytochrome bf complex in isolated chloroplasts.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>10632705</pmid><doi>10.1046/j.1432-1327.2000.01006.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | assembly Biological Transport Cell Membrane - chemistry Cell Membrane - metabolism chloroplast Chloroplasts - metabolism Cytochrome b Group - biosynthesis Cytochrome b Group - metabolism Cytochrome b6f Complex cytochrome bf complex Electron Transport Complex III Electrophoresis - methods Iron-Sulfur Proteins - genetics Iron-Sulfur Proteins - metabolism Mutation Pisum sativum Pisum sativum - metabolism Rieske FeS protein thylakoid membrane thylakoids Thylakoids - chemistry Thylakoids - metabolism Time Factors |
| title | Assembly of Rieske iron-sulphur protein into the cytochrome bf complex in thylakoid membranes of isolated pea chloroplasts |
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