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131.
Gong X Xie T Yu L Hesterberg M Scheide D Friedrich T Yu CA 《The Journal of biological chemistry》2003,278(28):25731-25737
An azido-ubiquinone derivative, 3-azido-2-methyl-5-methoxy[3H]-6-decyl-1,4-benzoquinone ([3H]azido-Q), was used to study the ubiquinone/protein interaction and to identify the ubiquinone-binding site in Escherichia coli NADH:ubiquinone oxidoreductase (complex I). The purified complex I showed no loss of activity after incubation with a 20-fold molar excess of [3H]azido-Q in the dark. Illumination of the incubated sample with long wavelength UV light for 10 min at 0 degrees C caused a 40% decrease of NADH:ubiquinone oxidoreductase activity. SDS-PAGE of the complex labeled with [3H]azido-Q followed by analysis of the radioactivity distribution among the subunits revealed that subunit NuoM was heavily labeled, suggesting that this protein houses the Q-binding site. When the [3H]azido-Q-labeled NuoM was purified from the labeled reductase by means of preparative SDS-PAGE, a 3-azido-2-methyl-5-methoxy-6-decyl-1,4-benzoquinone-linked peptide, with a retention time of 41.4 min, was obtained by high performance liquid chromatography of the protease K digest of the labeled subunit. This peptide had a partial NH2-terminal amino acid sequence of NH2-VMLIAILALV-, which corresponds to amino acid residues 184-193 of NuoM. The secondary structure prediction of NuoM using the Toppred hydropathy analysis showed that the Q-binding peptide overlaps with a proposed Q-binding motif located in the middle of the transmembrane helix 5 toward the cytoplasmic side of the membrane. Using the PHDhtm hydropathy plot, the labeled peptide is located in the transmembrane helix 4 toward the periplasmic side of the membrane. 相似文献
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135.
Ruan X Xu Q Mao HB Li GF Wei J Gong YD Kuang TY Zhao NM 《Journal of Protein Chemistry》2001,20(3):247-254
Changes in the protein secondary structure and electron transport activity of the Triton X-100-treated photosystem I (PSI) and photosystem II (PSII) complexes after strong illumination treatment were studied using Fourier transform-infrared (FT-IR) spectroscopy and an oxygen electrode. Short periods of photoinhibitory treatment led to obvious decreases in the rates of PSI-mediated electron transport activity and PSII-mediated oxygen evolution in the native or Triton-treated PSI and PSII complexes. In the native PSI and PSII complexes, the protein secondary structures had little changes after the photoinhibitory treatment. However, in both Triton-treated PSI and PSII complexes, short photoinhibition times caused significant loss of -helical content and increase of -sheet structure, similar to the conformational changes in samples of Triton-treated PSI and PSII complexes after long periods of dark incubation. Our results demonstrate that strong-light treatment to the Triton-treated PSI and PSII complexes accelerates destruction of the transmembrane structure of proteins in the two photosynthetic membranes. 相似文献
136.
Male accessory gland derived factors can stimulate oogenesis and enhance oviposition in Helicoverpa armigera (Lepidoptera: Noctuidae) 总被引:1,自引:0,他引:1
In Helicoverpa armigera, female moths began to lay eggs on the third day after emergence. Mating stimulated earlier egg maturation/oogenesis (P = 0.002) and oviposition (P < 0.01). We established a suitable bioassay model for the influence of male accessory glands (MAG) on the physiology of virgin females: Crude extracts of MAG (2- to 3-day-old) were injected into 2-day-old virgin females, and the injected females were dissected 20 h after mating. It was shown that crude extracts of MAG stimulated earlier egg maturation (P < 0.001) and oviposition (the oviposition ratio was more than 2 times the ratio of the control). Proteinaceous components in crude extracts purified by fractionation and sub-fractionation in reverse phase high performance liquid chromatography also stimulated earlier egg maturation (P < 0.01) and ovipositon (more than 2 times the ratio of the control), and we called them the oogenesis and ovipostion factors (OOSF). With SDS-PAGE, the molecular mass of the bands from OOSF was estimated to be between 55-66 KD. Arch. 相似文献
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138.
The nature of interaction between vaccinia virus and the surface of host cells as the first step in virus infection is undefined. A 32-kDa virus envelope protein has been identified as a cell surface binding protein (J.-S. Maa, J. F. Rodriguez, and M. Esteban, J. Biol. Chem. 265:1569-1577, 1990). To carry out studies on the structure-function relationship of this protein, the 32-kDa protein was obtained from Escherichia coli cells harboring the expression plasmid pT7Ek32. The recombinant polypeptide was found to have structural properties similar to those of the native virus envelope protein. Binding studies of 125I-labeled 32-kDa protein to cultured cells of various origins revealed that the E. coli-produced 32-kDa protein exhibited selectivity, specificity, and saturability. Scatchard analysis indicated about 4.5 x 10(4) sites per cell with a high affinity (Kd = 1.8 x 10(-9) M), suggesting interaction of the 32-kDa protein with a specific receptor. The availability of large quantities of the 32-kDa virus protein in bacteria will permit further structural and functional studies of this virus envelope protein and facilitate identification of the specific cell surface receptor. 相似文献
139.
Cd对桑叶品质,生理生化特性的影响及其机理研究 总被引:25,自引:2,他引:25
土壤中不同浓度的Cd对桑叶产量、品质、生理生化特性的影响以及Cd在桑叶亚细胞各组分中分布量的研究表明,当土壤Cd浓度小于22.3mg·kg-1时,桑叶产量、可溶性糖和含氯化合物含量高于或接近对照;桑叶叶绿素含量、细胞膜透性和超氧化物歧化酶、过氧化物酶及蛋白酶的活性无明显影响或有促进.当土壤Cd浓度大于22.3mg·kg-1时,Cd对桑叶产量、营养物质含量、生理生化作用的影响随之明显,并表现其毒害作用.根据Cd在桑叶亚细胞组分中的分布规律.对其影响机理进行了探讨,说明桑树是一种具有一定耐Cd性的经济作物.因此,可利用其耐Cd性,在被Cd污染土地上种植桑树. 相似文献
140.
Liu F Liang Z Shi J Yin D El-Akkad E Grundke-Iqbal I Iqbal K Gong CX 《FEBS letters》2006,580(26):6269-6274
Phosphorylation of tau protein is regulated by several kinases, especially glycogen synthase kinase 3beta (GSK-3beta), cyclin-dependent protein kinase 5 (cdk5) and cAMP-dependent protein kinase (PKA). Phosphorylation of tau by PKA primes it for phosphorylation by GSK-3beta, but the site-specific modulation of GSK-3beta-catalyzed tau phosphorylation by the prephosphorylation has not been well investigated. Here, we found that prephosphorylation by PKA promotes GSK-3beta-catalyzed tau phosphorylation at Thr181, Ser199, Ser202, Thr205, Thr217, Thr231, Ser396 and Ser422, but inhibits its phosphorylation at Thr212 and Ser404. In contrast, the prephosphorylation had no significant effect on its subsequent phosphorylation by cdk5 at Thr181, Ser199, Thr205, Thr231 and Ser422; inhibited it at Ser202, Thr212, Thr217 and Ser404; and slightly promoted it at Ser396. These studies reveal the nature of the inter-regulation of tau phosphorylation by the three major tau kinases. 相似文献