Purified PSI complexes from Spinacia Oleracea L. were exposed to the strong light (PFD=2300μmol m-2s-1) for various period. Along with the illumination the photo-damage process of pigments and proteins of PSI complex...Purified PSI complexes from Spinacia Oleracea L. were exposed to the strong light (PFD=2300μmol m-2s-1) for various period. Along with the illumination the photo-damage process of pigments and proteins of PSI complexes was investigated using absorption, fluorescence, circular dichroism (CD) spectroscopy and SDS-PAGE. It was found from the optical absorption spectra that the maximal ab-sorbance of PSI complexes decreased and maximal peaks blue-shifted during the illumination, and the forth derivative spectra demonstrated that the absorbance decreasing at red region mainly resulted from the aborbance decreasing of the long wavelength Chla, implying that the long-wavelength Chla was readily to be bleached. The CD signals contributed by LHCI decreased more rapidly than other CD signals con-tributed by Chla and Carotenoid, indicating that the LHCI was more sensitive to light than core complexes. It was observed by SDS-PAGE that some small polypeptides of PSI complexes were damaged earlier than reaction展开更多
Lhcb2 gene from pea (Pisum sativum L.) was subcloned into bacterial expression vector pET-3d, and its protein overexpressed was obtained from E. coli (BL21) containing PetpLhcb2 by site-directed mutagenesis method. Ba...Lhcb2 gene from pea (Pisum sativum L.) was subcloned into bacterial expression vector pET-3d, and its protein overexpressed was obtained from E. coli (BL21) containing PetpLhcb2 by site-directed mutagenesis method. Bacteria transformed with this construct yielded up to 40 percent of total protein of E. coli. Using the modified method with three subsequent cycles of freezing (1 min, -196℃) and thawing (15 min, 25℃), Lhcb2 protein purified was highly reconstituted with pigments to yield pigment-protein complexes. The reconstituted LHCB2 monomers were very similar to native LHCll monomers from spinach in partially denaturing polyacrylamide gel electrophoresis, fluorescence and absorbance spectroscopy. These results showed that Lhcb2 proteins overexpressed were reconstituted successfully with pigments and had similar organization and structure to the native LHCII monomers.展开更多
基金This work was supported by the National Natural Science Foundation of China (Grant No. 39890390) the State Key Basic Research and Development Program (Grant No. G1998010100) and the Innovative Foundation of Laboratory of Photosynthesis Basic Research
文摘Purified PSI complexes from Spinacia Oleracea L. were exposed to the strong light (PFD=2300μmol m-2s-1) for various period. Along with the illumination the photo-damage process of pigments and proteins of PSI complexes was investigated using absorption, fluorescence, circular dichroism (CD) spectroscopy and SDS-PAGE. It was found from the optical absorption spectra that the maximal ab-sorbance of PSI complexes decreased and maximal peaks blue-shifted during the illumination, and the forth derivative spectra demonstrated that the absorbance decreasing at red region mainly resulted from the aborbance decreasing of the long wavelength Chla, implying that the long-wavelength Chla was readily to be bleached. The CD signals contributed by LHCI decreased more rapidly than other CD signals con-tributed by Chla and Carotenoid, indicating that the LHCI was more sensitive to light than core complexes. It was observed by SDS-PAGE that some small polypeptides of PSI complexes were damaged earlier than reaction
文摘Lhcb2 gene from pea (Pisum sativum L.) was subcloned into bacterial expression vector pET-3d, and its protein overexpressed was obtained from E. coli (BL21) containing PetpLhcb2 by site-directed mutagenesis method. Bacteria transformed with this construct yielded up to 40 percent of total protein of E. coli. Using the modified method with three subsequent cycles of freezing (1 min, -196℃) and thawing (15 min, 25℃), Lhcb2 protein purified was highly reconstituted with pigments to yield pigment-protein complexes. The reconstituted LHCB2 monomers were very similar to native LHCll monomers from spinach in partially denaturing polyacrylamide gel electrophoresis, fluorescence and absorbance spectroscopy. These results showed that Lhcb2 proteins overexpressed were reconstituted successfully with pigments and had similar organization and structure to the native LHCII monomers.