Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components;...Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components; bands Ⅷ and Ⅸ (pI = 2.8-3.6) were chlorophyll-protein complexes. According to absorption and fluorescence spectra, band VII was designated as a novel phycocyanin-Chla/c2-protein complex (pI ≈ 3.4-3.7). These results indicated that phycocyanin is structurally and functionally coupled with chlorophyll-protein complex in C. placoidea, and probably interacted with electrostatic force in combination.展开更多
Natural phycobilisomes (PBSs) were isolated and purified from a red macroalga, Polysiphonia urceolata, by multi-step of sucrose gradient centrifugation, and were chemically stabilized by small molecule cross-linker ...Natural phycobilisomes (PBSs) were isolated and purified from a red macroalga, Polysiphonia urceolata, by multi-step of sucrose gradient centrifugation, and were chemically stabilized by small molecule cross-linker formaldehyde. The stabilized PBSs showed similar absorption and fluorescent properties at room temperature compared to natural PBSs and kept a steady F672/F580 value during more than 3 months of storage in 0.45 mol/L phosphate buffer (pH 6.8) or at low temperature at 77 K. The stabilized PBS migrated as a single band at mild PAGE and in 14-18 h of sucrose gradient centrifugation. All these characters indicated that the stabilized PBSs were stable, soluble, homogenous fluorescent particles with favorable spectroscopic features prepared under present conditions.展开更多
A new phycocyanin(PC) fluorescent subunit namedβ_2(18kDa) was isolated and characterized by both SDS-PAGE and isoelectric focusing(IEF) from a species of cryptophytic alga Chroomonas placoidea.PC was separated ...A new phycocyanin(PC) fluorescent subunit namedβ_2(18kDa) was isolated and characterized by both SDS-PAGE and isoelectric focusing(IEF) from a species of cryptophytic alga Chroomonas placoidea.PC was separated and purified by ammonium sulfate sedimentation followed by two steps of Sephadex G-100 chromatography.After denatured in 4 mol/L urea for 48 h,PC was divided into two fractions by passing through a Sephacryl S-100 chromatography column twice.The blue fraction(S-1) containedβsubunits with a maximal absorbance at 595 nm in visible light region.While the green fraction(S-2) enriched inαsubunits showed a characteristic long wavelength absorbance at 680-700 nm region and exhibited a relatively low molecular weight of 9.4(α_1) and 8.5 kDa(α_2).Fraction S-1 also consisted of two different fluorescent subunits with molecular weight of 20.1 kDa(β_1) and 18 kDa (β_2) and differed from each other on isoelectric points of pH 5.7(ft) and 6.0(ft),respectively.Further investigation of peptide sequence will help a lot in elucidating the new subunit ft that was smaller in size and more neutral than the known ft subunit,and may provide an alternative explanation in structure of cryptophytic phycobiliproteins.展开更多
文摘Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components; bands Ⅷ and Ⅸ (pI = 2.8-3.6) were chlorophyll-protein complexes. According to absorption and fluorescence spectra, band VII was designated as a novel phycocyanin-Chla/c2-protein complex (pI ≈ 3.4-3.7). These results indicated that phycocyanin is structurally and functionally coupled with chlorophyll-protein complex in C. placoidea, and probably interacted with electrostatic force in combination.
基金by Prize Fund for Excellent Middle Aged and Young Scientists of Shandong Province(No.2006BS02016)National Science Fund(No.30571720)
文摘Natural phycobilisomes (PBSs) were isolated and purified from a red macroalga, Polysiphonia urceolata, by multi-step of sucrose gradient centrifugation, and were chemically stabilized by small molecule cross-linker formaldehyde. The stabilized PBSs showed similar absorption and fluorescent properties at room temperature compared to natural PBSs and kept a steady F672/F580 value during more than 3 months of storage in 0.45 mol/L phosphate buffer (pH 6.8) or at low temperature at 77 K. The stabilized PBS migrated as a single band at mild PAGE and in 14-18 h of sucrose gradient centrifugation. All these characters indicated that the stabilized PBSs were stable, soluble, homogenous fluorescent particles with favorable spectroscopic features prepared under present conditions.
基金granted by the National Science Fund of China(No.40976083)
文摘A new phycocyanin(PC) fluorescent subunit namedβ_2(18kDa) was isolated and characterized by both SDS-PAGE and isoelectric focusing(IEF) from a species of cryptophytic alga Chroomonas placoidea.PC was separated and purified by ammonium sulfate sedimentation followed by two steps of Sephadex G-100 chromatography.After denatured in 4 mol/L urea for 48 h,PC was divided into two fractions by passing through a Sephacryl S-100 chromatography column twice.The blue fraction(S-1) containedβsubunits with a maximal absorbance at 595 nm in visible light region.While the green fraction(S-2) enriched inαsubunits showed a characteristic long wavelength absorbance at 680-700 nm region and exhibited a relatively low molecular weight of 9.4(α_1) and 8.5 kDa(α_2).Fraction S-1 also consisted of two different fluorescent subunits with molecular weight of 20.1 kDa(β_1) and 18 kDa (β_2) and differed from each other on isoelectric points of pH 5.7(ft) and 6.0(ft),respectively.Further investigation of peptide sequence will help a lot in elucidating the new subunit ft that was smaller in size and more neutral than the known ft subunit,and may provide an alternative explanation in structure of cryptophytic phycobiliproteins.