Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genom...Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genome-wide association analysis,the g-type lysozyme gene,which is named NaLyg in yellow drum(Nibea albiflora),was found to be a key candidate gene for disease resistance in response to Vibrio harveyi infection.The cDNA of NaLyg was 1025 bp,including four exons and three introns,and its open reading frame(ORF)had a full-length of 582 bp,encoding 193 amino acids.NaLyg was found to be conserved during evolution through bioinformatic analyses.The NaLyg protein possessed a sugar binding domain and three catalytic sites,including Glu71,Asp84 and Asp101.Quantitative qRT-PCR results confirmed that NaLyg gene mRNA was visibly increased after V.harveyi infection.The NaLyg protein purified by prokaryotic expression killed some gram-negative bacterial pathogens by inducing cell wall destruction,including V.harveyi,Aeromonas hydrophila and Edwardsiella tarda.Moreover,the NaLyg protein killed two gram-positive bacteria,Bacillus subtilis and Staphylococcus aureus.Taken together,the experimental results suggested that the NaLyg protein of N.albiflora played an important role in fighting bacterial infections.展开更多
Hydrophobic Val derivative Schiff base copper(II) complexes and dipeptide (AlaAla, GlyGly) derivative Schiff base copper(II) complexes were introduced into egg white lysozyme. X-ray crystal structure analysis revealed...Hydrophobic Val derivative Schiff base copper(II) complexes and dipeptide (AlaAla, GlyGly) derivative Schiff base copper(II) complexes were introduced into egg white lysozyme. X-ray crystal structure analysis revealed amino acid derivative Schiff base copper(II) complexes were obtained. Herein we discuss primarily on the binding mode of copper(II) of the complexes obtained with egg white lysozyme. The electron density of copper(II) ions was confirmed by X-ray crystal structure analysis. The Val derivative Schiff base copper(II) complex was weakly bound at Arg114 of egg white lysozyme. In other copper(II) complexes, binding of copper(II) ions with dissociated ligands to various residues was observed. The binding sites of copper(II) ions were compared with computational scientific predictions.展开更多
Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neu...Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neurological diseases.Herein,we applied Raman spectroscopy to study the Ni(II)ion effect on kinetics of amyloid fibrillation of hen egg white lysozyme(HEWL)in thermal and acidic conditions.Using the well-known Raman indicators for protein tertiary and secondary structures,we monitored and analyzed the concentration effect of Ni(II)ions on the unfolding of tertiary structures and the transformation of sec-ondary structures.The experimental evidence validates the accelerator role of the metal ion in the kinetics.Notably,the additional analysis of the amide I band profile,combined with thioflavin-T fluorescence assays,clearly indicates the inhibitory effect of Ni(II)ions on the formation of amyloid fibrils with organizedβ-sheets structures.Instead,a more significant promotion influence is affirmed on the assembly into other aggregates with disordered struc-tures.The present results provide rich information about the specific metal-mediated protein fibrillation.展开更多
In this study, we exhibited an amino acid (arginine and threonine) derivative Schiff base copper(II) complexes incorporating an azobenzene moiety as a photoresponsive site and conjugated it to egg white lysozyme, a we...In this study, we exhibited an amino acid (arginine and threonine) derivative Schiff base copper(II) complexes incorporating an azobenzene moiety as a photoresponsive site and conjugated it to egg white lysozyme, a well-known protein, to change ligand conformation under binding to lysozyme. Among several spectroscopic investigations, ESR clearly showed that the nitrogen atom of the amino acid residue of lysozyme was bound to the paramagnetic copper(II) ion of the complex, and UV light irradiation confirmed photoisomerization of the azobenzene moiety of the ligand to cis-form. The binding mode was considered by means of spectroscopic as well as computational methods, whereas complete crystallographic verification was still a preliminary stage.展开更多
G-protein coupled receptors （GPCRs） class C represent a distant group among the large family of GPCRs. This class includes the receptors for the main neurotransmitters, glutamate and gamma-aminobutyric acid （GABA）...G-protein coupled receptors （GPCRs） class C represent a distant group among the large family of GPCRs. This class includes the receptors for the main neurotransmitters, glutamate and gamma-aminobutyric acid （GABA）, and the receptors for Ca2＋, some taste and pheromone molecules, as well as some orphan receptors. Like any other GPCRs, these receptors possess a heptahelical domain （HD） involved in heterotrimeric G-protein activation, but most of them also have a large extracellular domain （VFT） responsible for agonist recognition and binding. These receptors are dimers, either homo or heterodimers. Then whereas have mGluRs is homodimers, GABAB receptor was the first heteromeric G-protein coupled receptor （GPCR） identified. Indeed, both GB1 and GB2 subunits appear necessary to get a functional GABAB receptor. We have demonstrated that the interactions be- tween VFT domain of both GB1 and GB2 were important for receptor activation. We have also shown the dynamic movement of trans-membrane of mGluRs within dimers. Then we have found that the GABAB receptor induced acti- vation of ERK1/2/CREB and protected neurons from apoptosis by trans-activating IGF-1R. We have also demon- strated that GABAB receptor activation has been modulated by the dynamic protein-protein interactions between re- ceptors and its downstream signal proteins such as FAK1 and Rap l. Finally, we have performed the HTS screening and found the first negative allosteric modulator for GABAB receptors.展开更多
Objective:To explore the nature of deficiency in spleen-yin syndrome,which could provide scientific theoretical support and practical guidance for clinical Traditional Chinese Medicine(TCM)syndrome differentiation bas...Objective:To explore the nature of deficiency in spleen-yin syndrome,which could provide scientific theoretical support and practical guidance for clinical Traditional Chinese Medicine(TCM)syndrome differentiation based on biology,and had a strong clinical significance.Methods:Serum Cu and Zn were detected by atomic absorption spectrophotometer,serum vitamin E by high performance liquid chromatography,serum vitamin C by 2,4-Dinitrophenylhydrazine Colorimetry,total superoxide dismutase(SOD)and Cu and Zn- SOD by the xanthine oxidase method,and malondialdehyde(MDA)by the 2-thiobarbituric acid method(TBA).Total antioxidant capacity was detected by a colorimetry kit.Amylase Activity was detected by an automatic biochemical analyzer.Lysozyme was detected by lysozyme detection plate,the diameter of bacteriolysis circle was measured and the corresponding content of lysozyme was obtained from a table of standard curve values.Results: No significant difference in total SOD and Cu,Zn-SOD was found between deficiency in spleen-yin group and normal group.However,such factors in deficiency in kidney-yin group were significantly lower than the other groups(P<0.05).The MDA content in both deficiency in spleen-yin group and deficiency in kidney-yin group were significantly higher than that of normal group(P<0.05),while the total antioxidant capacity was significantly lower than normal group(P<0.05).The vitamin E content in deficiency in kidney-yin group was significantly lower than that in the other two groups(P<0.05).No significant difference in the contents of vitamin C,Cu and Zn were observed in these groups.The Zn/Cu level in deficiency in kidney-yin group and the vitamin E level in deficiency in spleen-yin group decreased,but with no significant difference.Amylase activity in unit time in cases with deficiency in spleen-yin was lower than and had significant differences with that in normal cases,and higher than that in cases with deficiency in kidney-yin.The sectional velocity of saliva and the ratio of lysozyme in normal case group were significantly higher than other two groups,while deficiency in the spleen-yin group was significantly higher than the deficiency in kidney-yin group.Conclusion:All the results indicated that the objective pathological mechanism between the deficiency in spleen-yin and deficiency in kidney-yin was different.展开更多
基金supported by the National Natural Science Foundation of China(No.32072969)the National Key R&D Program of China(No.2022YFD2401002)+1 种基金the Natural Science Foundation of Fujian Province(No.2022 J01325)the Open Research Fund Program of Fujian Provincial Key Laboratory of Marine Fishery Resources and Eco-Environment(No.Z822280).
文摘Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genome-wide association analysis,the g-type lysozyme gene,which is named NaLyg in yellow drum(Nibea albiflora),was found to be a key candidate gene for disease resistance in response to Vibrio harveyi infection.The cDNA of NaLyg was 1025 bp,including four exons and three introns,and its open reading frame(ORF)had a full-length of 582 bp,encoding 193 amino acids.NaLyg was found to be conserved during evolution through bioinformatic analyses.The NaLyg protein possessed a sugar binding domain and three catalytic sites,including Glu71,Asp84 and Asp101.Quantitative qRT-PCR results confirmed that NaLyg gene mRNA was visibly increased after V.harveyi infection.The NaLyg protein purified by prokaryotic expression killed some gram-negative bacterial pathogens by inducing cell wall destruction,including V.harveyi,Aeromonas hydrophila and Edwardsiella tarda.Moreover,the NaLyg protein killed two gram-positive bacteria,Bacillus subtilis and Staphylococcus aureus.Taken together,the experimental results suggested that the NaLyg protein of N.albiflora played an important role in fighting bacterial infections.
文摘Hydrophobic Val derivative Schiff base copper(II) complexes and dipeptide (AlaAla, GlyGly) derivative Schiff base copper(II) complexes were introduced into egg white lysozyme. X-ray crystal structure analysis revealed amino acid derivative Schiff base copper(II) complexes were obtained. Herein we discuss primarily on the binding mode of copper(II) of the complexes obtained with egg white lysozyme. The electron density of copper(II) ions was confirmed by X-ray crystal structure analysis. The Val derivative Schiff base copper(II) complex was weakly bound at Arg114 of egg white lysozyme. In other copper(II) complexes, binding of copper(II) ions with dissociated ligands to various residues was observed. The binding sites of copper(II) ions were compared with computational scientific predictions.
基金supported by the National Natural Science Foundation of China(No.22073088,No.22027801 and No.21873089).
文摘Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neurological diseases.Herein,we applied Raman spectroscopy to study the Ni(II)ion effect on kinetics of amyloid fibrillation of hen egg white lysozyme(HEWL)in thermal and acidic conditions.Using the well-known Raman indicators for protein tertiary and secondary structures,we monitored and analyzed the concentration effect of Ni(II)ions on the unfolding of tertiary structures and the transformation of sec-ondary structures.The experimental evidence validates the accelerator role of the metal ion in the kinetics.Notably,the additional analysis of the amide I band profile,combined with thioflavin-T fluorescence assays,clearly indicates the inhibitory effect of Ni(II)ions on the formation of amyloid fibrils with organizedβ-sheets structures.Instead,a more significant promotion influence is affirmed on the assembly into other aggregates with disordered struc-tures.The present results provide rich information about the specific metal-mediated protein fibrillation.
文摘In this study, we exhibited an amino acid (arginine and threonine) derivative Schiff base copper(II) complexes incorporating an azobenzene moiety as a photoresponsive site and conjugated it to egg white lysozyme, a well-known protein, to change ligand conformation under binding to lysozyme. Among several spectroscopic investigations, ESR clearly showed that the nitrogen atom of the amino acid residue of lysozyme was bound to the paramagnetic copper(II) ion of the complex, and UV light irradiation confirmed photoisomerization of the azobenzene moiety of the ligand to cis-form. The binding mode was considered by means of spectroscopic as well as computational methods, whereas complete crystallographic verification was still a preliminary stage.
文摘G-protein coupled receptors （GPCRs） class C represent a distant group among the large family of GPCRs. This class includes the receptors for the main neurotransmitters, glutamate and gamma-aminobutyric acid （GABA）, and the receptors for Ca2＋, some taste and pheromone molecules, as well as some orphan receptors. Like any other GPCRs, these receptors possess a heptahelical domain （HD） involved in heterotrimeric G-protein activation, but most of them also have a large extracellular domain （VFT） responsible for agonist recognition and binding. These receptors are dimers, either homo or heterodimers. Then whereas have mGluRs is homodimers, GABAB receptor was the first heteromeric G-protein coupled receptor （GPCR） identified. Indeed, both GB1 and GB2 subunits appear necessary to get a functional GABAB receptor. We have demonstrated that the interactions be- tween VFT domain of both GB1 and GB2 were important for receptor activation. We have also shown the dynamic movement of trans-membrane of mGluRs within dimers. Then we have found that the GABAB receptor induced acti- vation of ERK1/2/CREB and protected neurons from apoptosis by trans-activating IGF-1R. We have also demon- strated that GABAB receptor activation has been modulated by the dynamic protein-protein interactions between re- ceptors and its downstream signal proteins such as FAK1 and Rap l. Finally, we have performed the HTS screening and found the first negative allosteric modulator for GABAB receptors.
文摘Objective:To explore the nature of deficiency in spleen-yin syndrome,which could provide scientific theoretical support and practical guidance for clinical Traditional Chinese Medicine(TCM)syndrome differentiation based on biology,and had a strong clinical significance.Methods:Serum Cu and Zn were detected by atomic absorption spectrophotometer,serum vitamin E by high performance liquid chromatography,serum vitamin C by 2,4-Dinitrophenylhydrazine Colorimetry,total superoxide dismutase(SOD)and Cu and Zn- SOD by the xanthine oxidase method,and malondialdehyde(MDA)by the 2-thiobarbituric acid method(TBA).Total antioxidant capacity was detected by a colorimetry kit.Amylase Activity was detected by an automatic biochemical analyzer.Lysozyme was detected by lysozyme detection plate,the diameter of bacteriolysis circle was measured and the corresponding content of lysozyme was obtained from a table of standard curve values.Results: No significant difference in total SOD and Cu,Zn-SOD was found between deficiency in spleen-yin group and normal group.However,such factors in deficiency in kidney-yin group were significantly lower than the other groups(P<0.05).The MDA content in both deficiency in spleen-yin group and deficiency in kidney-yin group were significantly higher than that of normal group(P<0.05),while the total antioxidant capacity was significantly lower than normal group(P<0.05).The vitamin E content in deficiency in kidney-yin group was significantly lower than that in the other two groups(P<0.05).No significant difference in the contents of vitamin C,Cu and Zn were observed in these groups.The Zn/Cu level in deficiency in kidney-yin group and the vitamin E level in deficiency in spleen-yin group decreased,but with no significant difference.Amylase activity in unit time in cases with deficiency in spleen-yin was lower than and had significant differences with that in normal cases,and higher than that in cases with deficiency in kidney-yin.The sectional velocity of saliva and the ratio of lysozyme in normal case group were significantly higher than other two groups,while deficiency in the spleen-yin group was significantly higher than the deficiency in kidney-yin group.Conclusion:All the results indicated that the objective pathological mechanism between the deficiency in spleen-yin and deficiency in kidney-yin was different.
