Artificial chaperone-assisted renaturation of R-PE subunits

Xie, Jie; Li, Donghui; Zhang, Jianping; Zhao, Jingquan

doi:10.1039/b209089a

Cited by 7 publications

(2 citation statements)

References 20 publications

(25 reference statements)

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“…R-PE was isolated and purified from Polysiphonia urceolata, a marine red alga collected on the seashore in Qingdao, Shandong province, China, by a procedure reported previously (Ma et al 1981;Xie et al 2003) and mainly in a hexameric state (Zeng et al 1985). R-PE purity and activity was characterized by the absorbance ratio at 565 to 280 nm (A565/A280) which was more than 5 in this work.…”

Section: Methodsmentioning

confidence: 99%

Ultrafast energy transfer pathways in R-phycoerythrin from Polysiphonia urceolata

et al. 2011

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Energy transfer (ET) processes between chromophores in R-phycoerythrin (R-PE) from Polysiphonia urceolata were studied by use of ultrafast spectroscopic methods. Several primary ET pathways were elaborated. A fluorescence decay component with a time constant of several hundred picoseconds observed by streak camera is tentatively assigned to the reversible formation of exciton traps between α84 and β84 pigment pairs. In order to investigate much faster ET processes in R-PE, a noncollinear optical parametric amplifier based femtosecond time-resolved transient fluorescence spectrometer was employed. The results reveal that the ET between α84 and β84 pigment pair has a time constant of 1-2 ps; the energy migration between α84 and β84 pairs within the R-PE trimer has a time constant of 30-40 ps. We also demonstrated an ET process from phycourobilin to phycoerythrobilin with a time constant as fast as 2.5-3.0 ps, which was directly observed in fluorescence kinetics by selective excitation of the phycourobilin molecules acting as the energy donor.

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Section: Methodsmentioning

confidence: 99%

Ultrafast energy transfer pathways in R-phycoerythrin from Polysiphonia urceolata

et al. 2011

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“…For molecular weight measurements, the renatured subunits derived by dialysis against redistilled water, avoiding any salt, were lyophilized and then determined on a Biflex III matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) (Bruker, Billerica, MA, USA) in the Beijing MS center, Institute of Chemistry, Chinese Academy of Science. 23 SDS-PAGE was performed in a vertical slab gel apparatus using discontinuous gels as described previously. 24 50 ll of Laemmli sample buffer was added into 50 ll R-PC (or subunits) and the mixture was heated for 10 min at 95 • C. 30 ll of the mixture were loaded on the 12% polyacrylamide slab gel containing 0.1% (w/v) SDS with a stacking gel of 4% polyacrylamide.…”

Section: Renaturation and Characteristics Of The Subunitsmentioning

confidence: 99%

Molecular properties of R-phycocyanin subunits from Polysiphonia urceolata in potassium phosphate buffer

Xie

Zhang

et al. 2008

Photochem Photobiol Sci

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The two subunits of R-phycocyanin from Polysiphonia urceolata were isolated and renatured. The renatured subunits were characterized by electrophoresis, molecular weights and spectra. The blue-shifted spectra, fluorescence recovery and restoring of the energy transfer suggested correct refolding of the subunits. The molecular properties of the subunits in potassium phosphate buffer (KPB) were investigated in detail. The total fluorescence yields (Q(T)) of the beta subunit declined while the energy transfer efficiency (E(T)) in the beta subunit was promoted with the increase of KPB concentration. On the other hand, both Q(T) and E(T) were enhanced with the increasing of the subunit concentrations. Based on the structural information, the fluorescence quenching in high concentrations of KPB was ascribed to less rigid chromophores caused by the weakening of the hydrogen-bond interaction network, while the enhancement of the fluorescence and E(T) was due to the aggregation of the subunits in the ionic solvent. Aggregation was confirmed by cysteine-assisted promotion of renaturation yield and stability, as well as equilibrium unfolding tests. Optimal conditions were proposed for the refolding/unfolding studies, under which the subunits were mainly monomeric. Compared to that in C-PC, the blue-shifted spectrum of PCB in R-PC is suggested to bring larger energy transfer efficiency, probably due to the necessity of the light harvesting for P. urceolata living in deep water.

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How CTAB assists the refolding of native and recombinant lysozyme

Wang

Lin

et al. 2005

Biochemical Engineering Journal

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Artificial chaperone-assisted renaturation of R-PE subunits

Cited by 7 publications

References 20 publications

Ultrafast energy transfer pathways in R-phycoerythrin from Polysiphonia urceolata

Ultrafast energy transfer pathways in R-phycoerythrin from Polysiphonia urceolata

Molecular properties of R-phycocyanin subunits from Polysiphonia urceolata in potassium phosphate buffer

How CTAB assists the refolding of native and recombinant lysozyme

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