The mitochondrial function was impaired in APP knockout mouse embryo fibroblast cells

The amyloid precursor protein (APP) is recognized as the source of Aβ, which plays an important role in Alzheimer’s disease. However, the biological function of APP is obscure. Previous studies showed that mitochondria could be a target of APP. In this work, APP knockout mouse embryo fibroblast (MEF) cells were used to test if APP plays any role in maintaining the mitochondrial function. As the result, APP knockout MEF cells (APP-/- cells) showed the abnormal mitochondrial function, including slower cell proliferation, lower mitochondrial membrane potential, lower intracellular ROS, higher mitochondrial membrane fluidity and lower cytochrome c oxidase activity than their wild-type counterparts. However, no change was found in the amount of mitochondria in MEF APP-/-cells.

Progress in artificial control system for gene expression

Along with the increasingly wide application of transgenic techniques, new stricter criteria have been raised for controlling the expression of exogenous genes. For these demands, a series of artificial control systems for gene expression have been developed and testified in recent years, which can control exogenous genes expression in exact time and certain level by administration of a specific drug or hormone. The successful construction of these systems offers a practicable method to control precise expression of exogenous gene in organisms, and raises the feasibility of wide application of gene therapy.

Endogenously generated amyloid β increases membrane fluidity in neural 2a cells

The effect of endogenously generated amyloid β on membrane fluidity was investigated in Neural 2a cells stably expressing Swedish mutant amyloid precursor protein (APPswe). Membrane fluidity was studied by fluorescence polarizability using 1,6-Diphenyl-1,3,5-Hexatriene (DPH) as the fluorescence probe. It was found that the membrane fluidity in APPswe cells was significantly higher than that in its wild type counterparts. Alleviating the effect of amyloid β either by γ secretase activity inhibition or by amyloid antibody treatment decreased membrane fluidity, which indicated an important role of amyloid β in increasing membrane fluidity. Treatment using amyloid β channel blocker, tromethamine and NA4 suggested that channel formed by amyloid β on the cell membrane is a way through which amyloid β takes its membrane fluidizing effect.

Effect of calcium ions on the secondary structures of photosystemⅡ and its relations with photoinhibition

Calcium ions play an important role in the oxygen_evolving process of photosystem Ⅱ as demonstrated in many experiments. The changes of the secondary structures of PS Ⅱ induced by the depletion of Ca 2+ were reported. The results indicated that the removal of Ca 2+ led to the transition of α helix to turns and sheet structures. While Ca 2+ was re_added to the media, only the structures changed to turns could be recovered. The protein conformational changes of PS Ⅱ during the donor side photoinhibition induced by the depletion of Ca 2+ were also studied. This showed that the protein conformational changes differed between the control and Ca 2+ _depleted samples in a short period of illumination (within 10 min). However, the changes became similar when the illumination time was increased.

The photodamage process of pigments and proteins of PSI complexes from Spinacia Oleracea L.

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

Simulation for the primary electronic donor of photosystem II in vitro

Histidine coordinated to Chl a is a distinct characteristic of Chl a in vivo. By usinghistidine analogue of 1-methylimidazole (C4H6N2) and measuring the UV/vis absorption, CD and MCD spectra of the interaction between C4H6N2and Chl a in CCI4, we have obtained that: (i) In pure CCI4 solvent, Chl a molecule is in five-coordinate state, and two Chl a molecules form an asymmetric compact-dimer with strong coupling interaction. We propose that the two Chl a molecules are connected by two unequally coordinated Mg-O bonds (the two oxygen atoms come from the C=O of C131 keto and C17 ester, respectively); (ii) when the molar ratio of C4H6N2/Chl a is 0.5 or 1 (corresponding to 2Chl a #1C4H6N2 and 2Chl a ·2C4H6N2, respectively), significant changes have been observed in the absorption, CD and MCD spectra, which indicate that the Chl a remains in dimer form, but the coupling interaction between them reduces greatly. We postulate that C4H6N2 replaces the ligation of C=O of C17 ester and C131 keto to Mg atoms

Preparation of the selenome-thionine derivative of tab-toxin resistance protein

In order to obtain phase information for the X-ray diffraction of tabtoxin resistance protein (TTR) crystal using the MAD phasing method, a selenomethionine (Se-Met) derivative of TTR was overexpressed in E. coli strain M15, with pQE-30 plasmid, through IPTG induction in M9 medium containing Se-Met. The product was purified to an estimated homogeneity of greater than 95% according to SDS-PAGE, by a Ni-NTA metal affinity followed by a Mono Q anion exchange column chromatography. The successful substitution of Se-Met for methionine (Met) was confirmed by MALDI-TOF and ESI-Quadrupole Mass Spectrometry analysis. The derivative crystal was obtained using similar conditions as those for the native.

Amyloid precursor protein-mediated modulation of capacitive calcium entry

Alzheimer's disease(AD) is a progressive neurodegenerative disorder.In the present study,the function of amyloid precursor protein(APP) in modulating capacitive calcium entry(CCE),a refilling mechanism for depleted intracellular calcium stores,was investigated.CCE in neural 2a(N2a) cells stably expressing wild-type human APP was lower than in wild-type N2a cells,while CCE in APP knockout mouse embryonic fibroblast(MEF) cells was higher than in their wild-type counterparts.We demonstrate that wild-type APP depresses CCE.Furthermore,using N2a cells transfected with C-terminal APP fragments,we show that these fragments anchored in the cell membrane play an important role in CCE depression.