It remains unknown whether a sucrose transporter mediates sugar signaling. Here, we report that the Arabidopsis （Arabidopsis thaliana） sucrose transporter SUT4 interacts with five members of the Arabidopsis cytochro...It remains unknown whether a sucrose transporter mediates sugar signaling. Here, we report that the Arabidopsis （Arabidopsis thaliana） sucrose transporter SUT4 interacts with five members of the Arabidopsis cytochrome b5 （Cyb5） family, and sucrose represses the interaction between SUT4 and a Cyb5 member Cyb5-2/A. We observed that down- regulation of SUT4 and three cytochrome b5 members （Cyb5-2, Cyb5-4, and Cyb5-6） confers the sucrose- and glucose- insensitive phenotypes in the sucrose/glucose-induced inhibition of seed germination. The sut4 cybS-2 double mutant displays slightly stronger sucrose/glucose-insensitive phenotypes than either the sut4 or cyb5-2 single mutant. We showed that the SUT4/Cyb5-2-mediated signaling in the sucrose/glucose-induced inhibition of seed germination does not require ABA or the currently known ABI2/ABI4/ABI5-mediated signaling pathway（s）. These data provide evidence that the sucrose transporter SUT4 interacts with Cyb5 to positively mediate sucrose and glucose signaling in the sucrose/ glucose-induced inhibition of seed germination.展开更多
The solution structure of bovine cytochrome b 5 mutant (E44A/E48A/E56A/D60A) has been obtained from 1H NMR spectra measured at 600 MHz. The final RMSD values with respect to the average structure are (0.056±0.009...The solution structure of bovine cytochrome b 5 mutant (E44A/E48A/E56A/D60A) has been obtained from 1H NMR spectra measured at 600 MHz. The final RMSD values with respect to the average structure are (0.056±0.009) nm and (0.105±0.010) nm for backbone and all heavy atoms, respectively. The solution structure, when compared with the X ray structure of wild type, has no significant changes in the whole folding and secondary structure. Small changes are restricted to local conformation near mutated sites. [WT5HZ]展开更多
Glu44, Glu48, Glu56 and Asp60 are the negatively charged residues located at the molecular surface of cytochrome b 5. Two mutants of cytochrome b 5 were prepared, in which two or all of these four residues were muta...Glu44, Glu48, Glu56 and Asp60 are the negatively charged residues located at the molecular surface of cytochrome b 5. Two mutants of cytochrome b 5 were prepared, in which two or all of these four residues were mutated to alanines. The mutations give rise to slightly positive shifts of the redox potentials of cytochrome b 5 and obvious decrease of the cytochrome b 5-cytochrome c binding constants and electron transfer rates. The crystal structures of the two mutants were determined at 0.18 nm resolution, showing no alteration in overall structures and exhibiting slight changes in the local conformations around the mutation sites as compared with the wild-type protein. Based on the crystal structure of the quadruple-site mutant, a model for the binding of this mutant with cytochrome c is proposed, which involves the salt bridges from Glu37, Glu38 and heme propionate of cytochrome b 5 to three lysines of cytochrome c and can well account for the properties and behaviors of this mutant.展开更多
To understand effect of π-stacking interactions between the side chain of aromatic amino acids and the por-phyrin ring on structures and properties in cytochrome b5 (cyt b5), the Phe58 residue was mutated to tyrosine...To understand effect of π-stacking interactions between the side chain of aromatic amino acids and the por-phyrin ring on structures and properties in cytochrome b5 (cyt b5), the Phe58 residue was mutated to tyrosine and tryptophan, respectively by site-directed mutagenesis. The denaturation of cyt bs F58W and F58Y toward guanidine hydrochloride was examined by UV-visible and fluorescence spectroscopy. The kinetics of heme transfer reactions between apo-myoglobin and the mutants were studied. The results indicated that the mutation of F58 residue for Y58 or W58 reduced the interaction between of peptide and the heme group, resulting in decrease of the Tm and Cm values of the proteins, increase of the heme transfer reaction rate, and shifts of the redox potential.展开更多
基金This research was supported by the National Natural Science Foundation of China
文摘It remains unknown whether a sucrose transporter mediates sugar signaling. Here, we report that the Arabidopsis （Arabidopsis thaliana） sucrose transporter SUT4 interacts with five members of the Arabidopsis cytochrome b5 （Cyb5） family, and sucrose represses the interaction between SUT4 and a Cyb5 member Cyb5-2/A. We observed that down- regulation of SUT4 and three cytochrome b5 members （Cyb5-2, Cyb5-4, and Cyb5-6） confers the sucrose- and glucose- insensitive phenotypes in the sucrose/glucose-induced inhibition of seed germination. The sut4 cybS-2 double mutant displays slightly stronger sucrose/glucose-insensitive phenotypes than either the sut4 or cyb5-2 single mutant. We showed that the SUT4/Cyb5-2-mediated signaling in the sucrose/glucose-induced inhibition of seed germination does not require ABA or the currently known ABI2/ABI4/ABI5-mediated signaling pathway（s）. These data provide evidence that the sucrose transporter SUT4 interacts with Cyb5 to positively mediate sucrose and glucose signaling in the sucrose/ glucose-induced inhibition of seed germination.
文摘The solution structure of bovine cytochrome b 5 mutant (E44A/E48A/E56A/D60A) has been obtained from 1H NMR spectra measured at 600 MHz. The final RMSD values with respect to the average structure are (0.056±0.009) nm and (0.105±0.010) nm for backbone and all heavy atoms, respectively. The solution structure, when compared with the X ray structure of wild type, has no significant changes in the whole folding and secondary structure. Small changes are restricted to local conformation near mutated sites. [WT5HZ]
文摘Glu44, Glu48, Glu56 and Asp60 are the negatively charged residues located at the molecular surface of cytochrome b 5. Two mutants of cytochrome b 5 were prepared, in which two or all of these four residues were mutated to alanines. The mutations give rise to slightly positive shifts of the redox potentials of cytochrome b 5 and obvious decrease of the cytochrome b 5-cytochrome c binding constants and electron transfer rates. The crystal structures of the two mutants were determined at 0.18 nm resolution, showing no alteration in overall structures and exhibiting slight changes in the local conformations around the mutation sites as compared with the wild-type protein. Based on the crystal structure of the quadruple-site mutant, a model for the binding of this mutant with cytochrome c is proposed, which involves the salt bridges from Glu37, Glu38 and heme propionate of cytochrome b 5 to three lysines of cytochrome c and can well account for the properties and behaviors of this mutant.
基金This work was supported by the National Natural Science Foundation of China (Grant No. 20071009).
文摘To understand effect of π-stacking interactions between the side chain of aromatic amino acids and the por-phyrin ring on structures and properties in cytochrome b5 (cyt b5), the Phe58 residue was mutated to tyrosine and tryptophan, respectively by site-directed mutagenesis. The denaturation of cyt bs F58W and F58Y toward guanidine hydrochloride was examined by UV-visible and fluorescence spectroscopy. The kinetics of heme transfer reactions between apo-myoglobin and the mutants were studied. The results indicated that the mutation of F58 residue for Y58 or W58 reduced the interaction between of peptide and the heme group, resulting in decrease of the Tm and Cm values of the proteins, increase of the heme transfer reaction rate, and shifts of the redox potential.
