Depletion of phosphatidyl-glycerol head-group induces changes in oxygen evolution and protein secondary struc-tures of photosystem Ⅱ

The techniques of oxygen electrode polarogra-phy and Fourier transform infrared (FT-IR) spectroscopy were employed to explore the roles of polar head-group of phosphatidylglycerol (PG) molecules in the functional and structural aspects of photosystem Ⅱ (PS Ⅱ) through enzymatic approach. It was shown that the depletion of PG by treatment of phospholipase C (PLC) on PS Ⅱ particles caused the inhibition of oxygen evolving activity in PS Ⅱ. This effect also gave rise to changes in the protein secondary structures of PS Ⅱ, that is, an increase in a-helical conformation which is compensated by the loss of p-strand structures. It revealed that the head-group of PG molecules plays an important structural role in the maintenance of normal structure of PS Ⅱ proteins, which is required to maintain the appropriate physiological activity of the PS Ⅱ complex such as the oxygen evolving activity. It is suggested that there most probably exist hydrogen-bonding interactions between PG molecules and PS Ⅱ proteins.     

Depletion of phosphatidylglycerol head-group induces changes in oxygen evolution and protein secondary structures of photosystemⅡ

The techniques of oxygen electrode polarography and Fourier transform infrared (FT-IR) spectroscopy were employed to explore the roles of polar head-group of phosphatidylglycerol (PG) molecules in the functional and structural aspects of photosystemⅡ(PSⅡ) through enzymatic approach. It was shown that the depletion of PG by treatment of phospholipase C (PLC) on PSⅡ particles caused the inhibition of oxygen evolving activity in PSⅡ. This effect also gave rise to changes in the protein secondary structures of PSⅡ, that is, an increase in α-helical conformation which is compensated by the loss of β-strand structures. It revealed that the head-group of PG molecules plays an important structural role in the maintenance of normal structure of PSⅡ proteins, which is required to maintain the appropriate physiological activity of the PSⅡ complex such as the oxygen evolving activity. It is suggested that there most probably exist hydrogen-bonding interactions between PG molecules and PSⅡ proteins.     

Low pH-induced conformational changes in 33 kD protein of photosystem Ⅱ

33 kD protein, located on the lumen side ofthylakoid membranes, is one of three extrinsic proteins ofphotosystem Ⅱ (PS Ⅱ ). Previous study showed that NBSmodification of W241, the only tryptophan in 33 kD protein,is helpful for understanding the function of W241 in main-taining functional conformation of 33 kD protein. In thispaper, studies of both circular dichroism and fluorescencespectra showed that upon decreasing pH from 6.2 to 2.5, theconformation of soluble 33 kD protein changed significantly,with an increase or a decrease in percentage of random coilor (z-helix and turns. The changes in secondary structures ofthis protein are pH reversible. After NBS modification at pH2.5, the conformational change of 33 kD protein was keptfixed. The CD ellipticity at 200 nm for NBS-modified 33 kDprotein is much lower than that for control, indicating that the unfolding degree of 33 kD protein was enhanced after the NBS modification. Moreover, the conformational flexibility islost in NBS-modified 33 kD protein, and the conformationalchange becomes pH irreversible, indicating that NBS modi-fication blocked the reversibility of conformational change of33 kD protein. The specific binding capability of NBS-modi-fled 33 kD protein is much lower than that of low pH-treatedcontrol. Furthermore, the rebinding of modified protein on PS Ⅱ membranes cannot restore the activity of oxygen evo-lution. We suggest that it is low pH but not NBS modificationof W241 that leads to the conformational change of 33 kDprotein from one functional to another non-functional state.The significant capability of proton transport of 33 kD pro-tein is discussed.     

Multiple intracellular signallings involved in regulation of on channels by GH releasing or inhibitory hormones in pituitary somatotropes

Influx of Ca2-via Ca2+ channels is the major step triggering exocytosis of pituitary somatotropes to release growth hormone (GH). Voltage-gated Ca2+ and K+ channels, the primary determinants of the influx of Ca2+ in somatotropes, are regulated by GH-releasing hornone (GHRH) and somatostatin (SRIF) through G protein-coupled signalling systems. Using whole-cell patch-clamp techniques, the changes of the Ca2+ and K+ currents in primary cultured somatotropes were recorded and signalling systems were studied using pharmacological reagents and intracellular dialysis of non-permeable molecules including antibodies and antisense oligonucleotides. GHRH increased both L-and T-types Ca2+ currents and decreased transient (I4) and delayed rectified (Ik) K+ currents. The increase in Ca2+ currents by GHRH was mediated by cAMP/protein kinase A system but the decrease in K+ currents required normal function of protein kinase C system. The GHRH-induced alteration of Ca2+ and K+ currents augments the influx of Ca2+ , leading to an increase in the [ Ca2+ ]I and the GH secretion. In contrary, a significant reduction in Ca2+ currents and increase in K currents were obtained in response to SRIF. The ion channel response to SRIF was demonstrated as a membrane delimited pathway and can be recorded by classic whole-cell configuration, Intracellular dialysis of anti-αi3 antibodies attenuated the increase in K + currents by SRIF whereas anti-αo antibodies blocked the reduction in the Ca2+ current by SRIF. Dialysis of antisense oligonucleotides specific for αo2 sub-units also attenuated the inhibition of SRIF on the Ca2+current. The Gi3 protein mediated the increase in K + currents and the Go2 protein mediated the reduction in the Ca2 +current by SRIF. The SRIF-induced alteration of Ca2 + and K + currents diminished the influx of Ca2+ , leading to a decrease in the [ Ca2+ ]I and the GH secretion. It is therefore concluded that multiple signalling systems are employed in the ion channel response to GHRH or SRIF in somatotropes, which leads to an increase or decrease in the GH secretion.     

Relationships of mRNA-protein secondary structures in the human β-globin gene HBB and four variants

Single nucleotide polymorphism is an interesting problem that can alter gene expression,recode amino acids and affect protein function.Protein structural changes have generally been attributed to amino acid replacements,and only a few research efforts have examined the effects of mRNA structural changes to the conformation of the corresponding protein coded by the mRNA.In the present study,the human β-globin HBB gene and four variants were examined.The mRNA secondary structures were constructed using the dynamic extended folding method and the encoded protein secondary structures were obtained from related databases.Comparisons were performed between these structures before and after mutations were introduced into the mature mRNAs and the proteins.We focused on the structural changes from mRNA to protein and found that regular protein conformations tend to match stable mRNA regions,whereas irregular protein conformations,such as β/γ turns and random coils,often match unstable mRNA regions.Mutations within unstable regions can alter the mRNA secondary structure and leave footprints in the protein structure.Comparison of the mRNA-protein secondary structure relationships represents a potential strategy to explore protein functional changes.     

Phosphatidylglycerol effect on oxygen-evolving activity in Ca2+-depleted photosystem Ⅱ

The effect of anionic phosphatidylglycerol (PG) on oxygen evolution in a photosystem Ⅱ (PSⅡ) particle depleted of Ca2+ (designated dCaPSⅡ) has been investigated. The major finding is the observation of a new role of PG in the PSⅡ function. That is, PG restores nearly the lost oxygen evolution in dCaPSⅡ particle owing to Ca2+ depletion to the levels in intact PSⅡ. Furthermore, there is a stimulation of oxygen-evolving activity in the dCaPSⅡ complexed with PG in the presence of exogenous CaCl2, which PG enhances increasingly oxygen evolution with increasing CaCl2 concentration. It is suggested that PG-induced oxygen evolution recovery of dCa PSⅡ particle results from resumption of normal structure in protein by PG effect, whereas the enhancement of oxygen evolution in complex subject to CaCl2 is ascribed to the optimization of such a structure due to coordination complex formation of Ca2+ ions with PG.     

Spectroscopic investigation of the interactions between gold nanoparticles and bovine serum albumin

The interactions between bovine serum albumin(BSA) and gold nanoparticles(AuNPs) ,and the conformational changes of BSA induced by this interaction,were investigated by UV-visible absorption spectroscopy,fluorescence spectroscopy,and Fourier transform infrared in combination with attenuated total reflection spectroscopy(ATR-FTIR) .The critical adsorption density for preventing AuNP aggregation in 0.1 mol/L phosphate buffered saline(pH 7.2) was 23 BSA molecules per gold particle or 3.8×1012 BSA molecules/cm2.BSA bound to the AuNPs with high affinity(binding constant Ks=7.59×108 L/mol) ,and the intrin-sic fluorescence of BSA was quenched by the AuNPs in accordance with the static quenching mechanism.Both fluorescence spectroscopy and ATR-FTIR showed that AuNPs induced conformational changes in BSA,which resulted in it becoming less compact and increased the polarity of the microenvironment around the tryptophan residue Trp-212.     

Mg~(2+)-induced LHC Ⅱ aggregation in thylakoid membrane

Mg2+-induced changes in 77K fluorescence spectra of wheat thylakoid membranes were analyzed by a Gaussian deconvolution program. All spectra were fitted well with 7 Gaussian sub-bands. The sub-band areas for the LHC Ⅱ macroaggregate (F699) and PS Ⅱ (F685 and F695) were increased and those for the LHC Ⅱ trimer (F681) and PS Ⅰ (F729 and F740) were decreased by Mg2+ . Moreover, it was found that the sub-band area ratio for F699 over F681 was dramatically enhanced by Mg2+ by 1.72 times. It was concluded that the LHC Ⅱ trimer in thylakoid membranes could be aggregated and transformed into its macroaggregate by Mg2+ on a large scale. The possible implication of the Mg2+-induced LHC Ⅱ aggregation was also discussed .     

Activity and Structure Changes of Arginine Kinase from Shrimp Feneropenaeus chinensis Muscle in Trifluoroethanol Solutions

Trifluoroethanol has often been used in protein folding studies. The changes in activity and unfolding of arginine kinase from shrimp Feneropenaeus Ghinensis muscle during denaturation in different concentrations of trifuoroethanol were investigated using far-ultraviolet circular dichroism and fluorescence emission spectra. Arginine kinase was inactivated in trifluoroethanol solutions. The tertiary and secondary structures of arginine kinase were also destroyed in the trifluoroethanol solutions. The unfolding and inactivation courses were measured and compared. Inactivation occurred prior to unfolding, which suggests that the arginine kinase active site is more easily damaged by the denaturant than the enzyme as a whole. The result also indicates that the arginine kinase active site is situated in a limited and flexible region of the enzymemolecule.     

Comparison of the Photosynthetic Characteristics of Two Developmental Stages in Nostoc sphaeroides Kutzing （Cyanophyta）

The photosynthetic activities between two main developmental stages, colony and hormogonium, of the edible cyanobacterium Nostoc sphaeroides Kutzing, were compared. Hormogonia have a higher content of chlorophyll than that of colonies. It showed that the ratios of phycocyain （PC）, allophycocyain （APC） and phycoerythrocyanin （PEC） in hormogonia and colonies were different. The room temperature chlorophyll fluorescence, 77 K chlorophyll fluorescence, measurements of PSⅠand PS Ⅱ activities all showed that colony has higher photosynthetic competence than hormogonia. Hormogonia had a higher respiration rate than colony, while their maximum photosynthetic oxygen evolution rates were very close. The responses of hormogonia and colonies to high light illuminations also were different. Both of their oxygen evolution rates decreased quickly with the prolonged high light illumination, but hormogonia can keep relatively higher PSⅡ activity （Fv/Fm） than that of colonies. The results suggested that colony was photosynthetically more competent than hormogonia, while the ability of hormogonia to tolerate high light illumination was higher than that of colony.     

Effect of Thrombin on the Apoptosis of Hippocampal Neurons in vitro

Hippocampal neurons were treated by thrombin and thrombin receptor activating peptides (TRAP). Cell survival rate was decreased in a dose-dependent manner by MTT assay. The numbers of apoptotic cell and apoptotic rate of hippocampal neurons treated by different concentrations of thrombin were increased in a dose-dependent manner by terminal deoxynucleotidyl transferase (TdT) mediated dUTP-biotin nick end-labeling (TUNEL) method and Flow Cytometry. When the concentration of thrombin is 40 U/mL, TUNEL positive cells and apoptotic rate of hippocampal neurons reached peak value, were 27. 3 4.0 and (29.333 4.633 ) %, respectively. Immunocytochemistry assay show that Bcl-2 protein expression was down- regulated and Bax protein expression was up-regulated with the concentration of thrombin increased. TRAP can mimic the effect of thrombin to induce apoptosis on hippocampal neurons. These data demonstrated that thrombin induced hippocampal neuron apoptosis in a dose-dependent manner through activating protease-activated protein-1 (PAR-1). The change in expression of Bcl-2 and Bax was related with the effect of high concentration thrombin induced apoptosis on hippocampal neurons.     

Increase in electron transfer activity in photosystem Ⅱ of spinach thylakoids caused by conversion of phosphatidyl- glycerol to phosphatidic acid molecules

The techniques of oxygen electrode polarography, sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE) and thin layer chromatography (TLC) were employed to investigate the effect of phospholipase D treatment on physiological function of spinach thylakoids. It was shown that the phospholipase D treatment on thylakoid resuited in the degradation of phosphatidylglycerol (PG) and occurrence of phosphatidic acid (PA). The changes of PG to PA molecules caused an increase in oxygen evolution in photosystem Ⅱ (PS Ⅱ), which was accompanied by an uncoupling effect on thylakoid membrane. It was revealed that the head-groups of PG molecules play an important role in themaintenance of the appropriate physiological activity of thylakoid membrane.     

Cu（Ⅱ） effect on the conformation of regenerated silk fibroin in dilute aqueous solution

Much attention has been paid to the natural mechanism of silkworm spinning due to the impressive mechanical properties of the natural fibers. In this work, we studied the effect of Cu（Ⅱ） ions on the secondary structure of Bombyx mori regenerated silk fibroin （SF） in dilute solution by circular dichroism （CD）. The results indicate that a given amount of Cu（Ⅱ） induces the SF conformational transition from random coil to β-sheet, however, further addition of Cu（Ⅱ） is unfavorable for this conversion. Meanwhile, the conformational changes induced by Cu（Ⅱ） follow a nucleation-dependent aggregation mechanism, which is similar to that found in Prion protein （PrP） denaturation and Aβ-peptide aggregations, leading to the neurodegenerative disease. This work would help one understand further the natural spinning process of silkworm. Additionally, it would be significant for the study of the nervous system diseases, because silk fibroin, extracted in large amounts from Bombyx mori silkworm gland, could be a proper model to study PrP denaturation and Aβ-peptide aggregations.     

Oscillatory change of SR-protein kinase activities during oocyte maturation meiosis in fish

The SR-protein kinase activity was analyzed and the cytological changes were observed during oocyte maturation in bisexual transparent color crucian carp ( Carassius auratus color variety) . The results revealed that the SR-protein kinase activity was sensitive to the artificially induced spawning hormones, and the change of oscillatory activity was similar to that of the maturation-promoting factor (MPF) kinase that regulates meiotic cell cycle in fish.     

Hypoosmotic shock induces a stateⅠtransition of photosynthetic apparatus in Dunaliella salina

The effects of hypoosmotic shock on state transition in darkness in Dunaliella salina were studied. When the concentration of NaCI in culture medium was dropped from 1.5 to 0.5 mol/L abruptly, the photosynthetic rate of D.salina declined, but the respiratory rate and intracellular ATP content increased in the dark. The FpsⅡ/FpsⅠ ratio at 77 K of D. salina cells exposed to hypoosmotic shock was higher than that of control cells, indicating that more excitation energy was distributed to PS Ⅱ in stressed D.salina cells upon illumination. A decrease in LHC Ⅱ phosphorylation level was also observed when D.salina was exposed to hypoosmotic shock. Thus a state Ⅰ transition of photosynthetic apparatus occurs when D.salina suffers hypoosmotic shock in darkness, which is supposed to be related to an enhancement of respiration and an increase in ATP content in stressed D.salina cells.     

A study on poly (N-vinyl-2-pyrrolidone) covalently bonded NiTi surface for inhibiting protein adsorption

Near equiatomic NiTi alloys have been extensively applied as biomaterials owing to its unique shape memory effect, superelasticity and biocompatibility. It has been demonstrated that surfaces capable of preventing plasma protein adsorption could reduce the reactivity of biomaterials with human blood. This motivated a lot of researches on the surface modification of NiTi alloy. In the present work, following heat and alkaline treatment and silanization by trichlorovinylsilane (TCVS), coating of poly (N-vinyl-2-pyrrolidone) (PVP) was produced on the NiTi alloy by gamma ray induced chemical bonding. The structures and properties of modified NiTi were characterized and in vitro biocompatibility of plasma protein adsorption was investigated. The results indicated that heat treatment at 823 K for 1 h could result in the formation of a protective TiO2 layer with “Ni-free” zone on NiTi surface. It was found that PVP was covalently bonded on NiTi surface to create a hydrophilic layer for inhibiting protein adsorption on the surface. The present work offers a green approach to introduce a bioorganic surface on metal and other polymeric or inorganic substrates by gamma irradiation.     

Influence of Dispersion of Nano-Zn0 Particles in Polymer Matrices on Properties of Relevant Nano Composite Fibers

The surface-passivated and non-surface-passivated zinc oxide nano-particles （marked as s-nanoZnO and ns-nanoZnO respectively） were evenly dispersed in polymer solutions with the aid of ultrasonic vibration to prepare nanocomposite film by free casting and to prepare nanocomposite fibers by wet spinning and to prepare nancomposites coating by surface smearing. The dispersion of s-nanoZnO and nsnanoZnO in PAN matrix were observed by transmittance electron microscopy, the mechanical properties of the relevant compesite samples were studied by INSRTON tensile strength tester. It was found that s-nanoZnO behaves a well-dispersed morphology in PAN films and fibers when its concentration was 2 wt% but ns-nanoZnO nano particles agglomerate into larger congeries in PAN films. It means that the surface-passivated process oft zinc oxide nano. particles was effective to disperse. The relative intensity and elonsation at break of s-nanoZnO-PAN composite fibers show maximum values with the increase of nano particle content in compesites （from 0 wt% to 2 wt% of s- nanoZnO）. The elasticity of the composite fibers increases whereas their modulus declines. Balanced the changes of the properties mentioned above, 2 wt% s-nanoZnO in PAN matrix is a proper content for the composite fibers spun by wet spinning. The result of surface smearing test means that the reactim between s-nanoZnO and polymer can be indicated by the color of nanocomposite surface coat on fibers.     

Distribution of lanthanum among the chloroplast subcomponents of spinach and its biological effects on photosynthesis： location of the lanthanum binding sites in photosystem Ⅱ

The effects of lanthanum at different concentrations on the related photosynthetic activities of Hill reaction, Mg^2＋-ATPase and Ca^2＋-ATPase in spinach chloroplast were studied. Experimental results showed that lanthanum can increase all the activities at suitable concentration （15-30 mg· L^-1）, however, it behaves toxically on them when over used （60 mg. L^-1）. To get an improved understanding of the mechanism of lanthanum effects on the photosynthesis of spinach, the different subcomponents in the chloroplast of the cultured spinach were isolated, and the content of lanthanum in each subcomponent was determined by ICP-MS. The results obtained indicated that among these different subcomponents, about 90% out of the total chloroplast lanthanum was located in photosystem Ⅱ （PS Ⅱ） while there was little lanthanum in photosystem Ⅰ （PS Ⅰ）. Moreover, size exclusion high performance liquid chromatography （SE-HPLC） coupled with online UV and ICP-MS detections was novelly used for locating lanthanum binding sites in PS Ⅱ proteins for the first time. It was found that lanthanum has two binding sites in PS Ⅱ： La associates with chlorophyll together with magnesium in PS Ⅱ by partly replacing magnesium and also shares the common binding sites of PS Ⅱ proteins together with the inorganic cofactors of calcium and manganese, influencing the process of photosynthesis.     

Cardiac protective role of a novel erythrocyte-derived depressing factor on rats and its Ca^2＋ mechanism

The cardiac protective role of a novel erythro-cyte-derived depressing factor (EDDF) on spontaneous hy-pertensive rats (SHR), calcium overload (CaO) rats and Wistar rats and its mechanism was evaluated. Mean artery pressure (MAP), heart rate (HR) and LVdp/dtmax were measured by physiological recorder. The effect of EDDF on the Ca2+-ATPase activity in myocardial sarcoplasmic reticu-lum (SR) of CaO rats was determined by inorganic phos-phate assay. Calcium transport in myocytes was measured by 45Ca2+ radioactive isotope measurement. The phosphoryla-tion levels of extracellular signal-regulated protein kinases (ERK1/2) in myocardial tissue of SHR and CaO rats were measured by Western blot method. And the ultrastructures of cardiac muscle cells were observed with the transmission electron microscope. The results indicated that EDDF could significantly decrease MAP, HR and LVdp/dtmax in a dose dependent manner (P < 0.05). It seems that the mechanism might relate with activating the Ca2+-APTase, enhancing the uptake and release of Ca2+ from SR (P < 0.05), decreasing the phosphorylation levels of ERK1/2 of myocytes (P < 0.01) and lightening the ultrastructural lesion of cardiac muscle cells. In CaO rats, the Ca2+-ATPase activity decreased clearly com-pared to control (64.99 7.16 vs 94.48 7.68 nmol·min-1 ·mg-1 protein, P < 0.01), while EDDF (100 mg/mL) could significantly increase the activity (87.93 ?9.54 vs 64.99 ?7.16, P < 0.05, n = 7). Both uptake and release rate of Ca2+ (祄ol 45Ca2+/g protein/min) from myocardial SR of CaO rats re-markably decreased compared to control (32.40 ?2.70 and 15.46 ?1.49 vs 61.09 ?10.89 and 25.47 ?4.29, P < 0.05); EDDF (100 mg/mL) could significantly stimulate their activi-ties (50.48 6.76 and 21.76 2.75 vs 32.40 2.70 and 15.46 1.49, P < 0.05). EDDF could evidently down-regulate the phosphorylation of ERK1/2 in myocardial tissue from SHR and CaO rats (P < 0.01), lighten the ultrastructural lesion of cardiac muscle cells of SHR as well. It is concluded that EDDF seems to play protective roles on both structure and function of heart, which closely related with amelioration of Ca2+ transport and inhibition of Ca2+-MAP kinase pathway.     

Preparation and Structural Characterization of Polystyrene-Rectorite Nanocornposites

The polystyrene/reetorite nanocomposites were prepared by free radical polymerization of styrene containing dispersed organophilic rectorite. The structures and thermal properties of these hybrids have been investigated by X-ray diffraction (XRD), fourier transform infrared (FT-IR), positron annihilation spectroscopy (PAS) and thermal gmvimetric analysis (TGA) techniques. It was found that exfoliation of reetorite in polystyrene (PS) matrix was achieved. The average free-volume radius in the PS/clay nanocomposites is generally same as that in PS. Along with increment of reetorite contents, the interface between reetorite and polystyrene matrix increases, and the free-volume concentration decreases obviously. And the polystyrene nanocomposites have higher thermal deeomrosition temoemture than oure PS.