Pub Date : 2017-01-01DOI: 10.1080/23312025.2017.1291877
Sarah M. Meeuwsen, An Nam Hodac, Lauren M. Adams, Ryan D. McMunn, Maxwell S. Anschutz, Kari J. Carothers, Rachel E. Egdorf, Peter M. Hanneman, Jonathan P. Kitzrow, Cynthia K. Keonigsberg, Óscar López-Martínez, P. A. Matthew, Ethan H. Richter, Jonathan E. Schenk, Heidi L. Schmit, M. Scott, Eva M. Volenec, S. Hati
Abstract Intrinsic dynamics of proteins are known to play important roles in their function. In particular, collective dynamics of a protein, which are defined by the protein’s overall architecture, are important in promoting the active site conformation that favors substrate binding and effective catalysis. The primary sequence of a protein, which determines its three-dimensional structure, encodes unique dynamics. The intrinsic dynamics of a protein actually link protein structure to its function. In the present study, coarse-grained normal mode analysis was performed to examine the intrinsic dynamic patterns of 24 different enzymes involved in primary metabolic pathways. We observed that each metabolic enzyme exhibits unique patterns of motions, which are conserved across multiple species and functionally relevant. Dynamic cross-correlation matrices (DCCMs) are visibly identical for a given enzyme family but significantly different from DCCMs of other protein families, reinforcing that proteins with similar function exhibit a similar pattern of motions. The present work also reasserted that correct identification of unknown proteins is possible based on their intrinsic mobility patterns.
{"title":"Investigation of intrinsic dynamics of enzymes involved in metabolic pathways using coarse-grained normal mode analysis","authors":"Sarah M. Meeuwsen, An Nam Hodac, Lauren M. Adams, Ryan D. McMunn, Maxwell S. Anschutz, Kari J. Carothers, Rachel E. Egdorf, Peter M. Hanneman, Jonathan P. Kitzrow, Cynthia K. Keonigsberg, Óscar López-Martínez, P. A. Matthew, Ethan H. Richter, Jonathan E. Schenk, Heidi L. Schmit, M. Scott, Eva M. Volenec, S. Hati","doi":"10.1080/23312025.2017.1291877","DOIUrl":"https://doi.org/10.1080/23312025.2017.1291877","url":null,"abstract":"Abstract Intrinsic dynamics of proteins are known to play important roles in their function. In particular, collective dynamics of a protein, which are defined by the protein’s overall architecture, are important in promoting the active site conformation that favors substrate binding and effective catalysis. The primary sequence of a protein, which determines its three-dimensional structure, encodes unique dynamics. The intrinsic dynamics of a protein actually link protein structure to its function. In the present study, coarse-grained normal mode analysis was performed to examine the intrinsic dynamic patterns of 24 different enzymes involved in primary metabolic pathways. We observed that each metabolic enzyme exhibits unique patterns of motions, which are conserved across multiple species and functionally relevant. Dynamic cross-correlation matrices (DCCMs) are visibly identical for a given enzyme family but significantly different from DCCMs of other protein families, reinforcing that proteins with similar function exhibit a similar pattern of motions. The present work also reasserted that correct identification of unknown proteins is possible based on their intrinsic mobility patterns.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2017.1291877","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48277448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1267421
H. Pham, L. Hoang, V. Phung
Abstract This study aims to investigate the hepatoprotective activity of Ganoderma lucidium mushroom growing on the dead ironwood tree in the Central Highlands of Vietnam. The total extract was orally administrated to experimental mice with hepatotoxicity induced by cyclophosphamide at the dose of 150 mg/kg (intraperitoneal injection). The liver malondialdehyde (MDA) content and the level of endogenous antioxidant glutathione (GSH) were measured. The results revealed that the total extract at the oral doses of 330, 230, and 120 mg/kg body weight which were equivalent to 5, 10, and 15 g/kg of dry material alleviated the increase of heaptic MDA content and restored the decrease of GSH level significantly which almost have the same potent as reference drug (silymarin). Biochemical observations were also supported with histopathological examination of liver. This finding demonstrated that G. lucidium in Vietnam could represent a promising approach for effective liver protective agents.
{"title":"Hepatoprotective activity of Ganoderma lucidium (Curtis) P. Karst against cyclophosphamide-induced liver injury in mice","authors":"H. Pham, L. Hoang, V. Phung","doi":"10.1080/23312025.2016.1267421","DOIUrl":"https://doi.org/10.1080/23312025.2016.1267421","url":null,"abstract":"Abstract This study aims to investigate the hepatoprotective activity of Ganoderma lucidium mushroom growing on the dead ironwood tree in the Central Highlands of Vietnam. The total extract was orally administrated to experimental mice with hepatotoxicity induced by cyclophosphamide at the dose of 150 mg/kg (intraperitoneal injection). The liver malondialdehyde (MDA) content and the level of endogenous antioxidant glutathione (GSH) were measured. The results revealed that the total extract at the oral doses of 330, 230, and 120 mg/kg body weight which were equivalent to 5, 10, and 15 g/kg of dry material alleviated the increase of heaptic MDA content and restored the decrease of GSH level significantly which almost have the same potent as reference drug (silymarin). Biochemical observations were also supported with histopathological examination of liver. This finding demonstrated that G. lucidium in Vietnam could represent a promising approach for effective liver protective agents.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1267421","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1236431
Amina Ghalem, I. Barbosa, R. Bouhraoua, Augusta Costa
Abstract In the southwestern Mediterranean Basin, cork oaks (Quercus suber L.) are periodically harvested for their cork. This natural product is valued by its homogeneity which heightens the importance of characterizing cork tissue discontinuities, or cork pores. Cork porosity profile in natural cork planks has been reported to be affected by forest management practices but, so far, has been scarcely addressed. We characterize the cork porosity profile in two contrasting cork oak woodland; at a mountain forest, in Western Algeria (absence of forest management) and at a peneplain “montado,” in southern Portugal (intensively managed toward the optimization of cork production). Image analysis techniques were applied on transverse sections of more than 40 cork samples from both woodland, and a stepwise discriminant analysis was used to discriminate between the cork pore features data-sets. Cork porosity profiles were similar between regions but; in the cork samples from Algeria, cork pores were having higher values for linear dimensions of pores (length and perimeter) and contrasting shape values (roundness) which depreciate cork quality, when compared to the cork samples from Portugal. However, improved woodland management strategies at Algeria should ensure adequate cork homogeneity and suitability for more valuable cork products.
{"title":"Comparing cork quality from Hafir-Zarieffet mountain forest (Tlemcen, Algeria) vs. Tagus basin Montado (Benavente, Portugal)","authors":"Amina Ghalem, I. Barbosa, R. Bouhraoua, Augusta Costa","doi":"10.1080/23312025.2016.1236431","DOIUrl":"https://doi.org/10.1080/23312025.2016.1236431","url":null,"abstract":"Abstract In the southwestern Mediterranean Basin, cork oaks (Quercus suber L.) are periodically harvested for their cork. This natural product is valued by its homogeneity which heightens the importance of characterizing cork tissue discontinuities, or cork pores. Cork porosity profile in natural cork planks has been reported to be affected by forest management practices but, so far, has been scarcely addressed. We characterize the cork porosity profile in two contrasting cork oak woodland; at a mountain forest, in Western Algeria (absence of forest management) and at a peneplain “montado,” in southern Portugal (intensively managed toward the optimization of cork production). Image analysis techniques were applied on transverse sections of more than 40 cork samples from both woodland, and a stepwise discriminant analysis was used to discriminate between the cork pore features data-sets. Cork porosity profiles were similar between regions but; in the cork samples from Algeria, cork pores were having higher values for linear dimensions of pores (length and perimeter) and contrasting shape values (roundness) which depreciate cork quality, when compared to the cork samples from Portugal. However, improved woodland management strategies at Algeria should ensure adequate cork homogeneity and suitability for more valuable cork products.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1236431","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1264162
P. Anushree, R. Naik, R. Satbhai, A. Gaikwad, C. Nimbalkar, J. Bahler
Abstract The disease-free (control) and blast infected leaf samples of 11 rice genotypes were evaluated for activity profile of defense-related and antioxidative enzymes. The amplification genomic DNA with two SSR markers RM124 and RM224 were also performed for identification of blast resistance and susceptible genotypes. The activity of chitinase, PAL and β-glucosidase of post pathogen-infected leaf samples increased significantly in all rice genotypes, thought the increase was comparable less in to blast susceptible genotypes Chimansal and EK-70. The activity of antioxidative enzymes was comparatively higher in the infected leaf of blast resistant genotypes recording highest increase in NLR-20104 and KJT-5. The activity of defense-related and antioxidative enzymes in the disease-free leaf samples differed among the genotypes and was even higher in the two blast susceptible genotypes. RM144 and RM224 SSR primers clearly amplified in blast resistant KJT-5, NLR-20104, KJT-2, Tetep genotypes whereas RM144 missing in susceptible Chimansal but prominently present in susceptible genotype EK-70. This study revealed that higher level of induction of defense-related and antioxidative enzymes and presence of specific amplified fragments with RM144 and RM224 could be useful for screening the resistant and susceptible rice genotypes against Magnaporthe oryzae.
{"title":"Differential biochemical response of rice (Oryza sativa L.) genotypes against rice blast (Magnaporthe oryzae)","authors":"P. Anushree, R. Naik, R. Satbhai, A. Gaikwad, C. Nimbalkar, J. Bahler","doi":"10.1080/23312025.2016.1264162","DOIUrl":"https://doi.org/10.1080/23312025.2016.1264162","url":null,"abstract":"Abstract The disease-free (control) and blast infected leaf samples of 11 rice genotypes were evaluated for activity profile of defense-related and antioxidative enzymes. The amplification genomic DNA with two SSR markers RM124 and RM224 were also performed for identification of blast resistance and susceptible genotypes. The activity of chitinase, PAL and β-glucosidase of post pathogen-infected leaf samples increased significantly in all rice genotypes, thought the increase was comparable less in to blast susceptible genotypes Chimansal and EK-70. The activity of antioxidative enzymes was comparatively higher in the infected leaf of blast resistant genotypes recording highest increase in NLR-20104 and KJT-5. The activity of defense-related and antioxidative enzymes in the disease-free leaf samples differed among the genotypes and was even higher in the two blast susceptible genotypes. RM144 and RM224 SSR primers clearly amplified in blast resistant KJT-5, NLR-20104, KJT-2, Tetep genotypes whereas RM144 missing in susceptible Chimansal but prominently present in susceptible genotype EK-70. This study revealed that higher level of induction of defense-related and antioxidative enzymes and presence of specific amplified fragments with RM144 and RM224 could be useful for screening the resistant and susceptible rice genotypes against Magnaporthe oryzae.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1264162","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1275403
T. Kwape, R. Majinda, P. Chaturvedi
Abstract The total phenol content, antioxidant and antidiabetic activity of Myrothamnus flabellifolius (MF) were investigated. The study revealed 70% ethanol/water (EW70), 70% methanol/water (MW70) and methanol (Me) extracts to contain the highest phenol content, 350 mg GAE/g, 300 mg GAE/g and 250 mg GAE/g, respectively (GAE/g—Gallic Acid Equivalent per gram). MW70 exhibited the highest radical [diphenyl picryl hydrazine (DPPH) and 2, 2-Azobis-3-ethyl benzthiazoline-6-sulphonic acid (ABTS+)] scavenging power. About 70% ethanol/water exhibited the highest reducing power, hydrogen peroxide and nitric oxide scavenging activities. The results of antidiabetic activity showed EW70 to possess the highest α-amylase inhibitory effect. The α-glucosidase activity was inhibited by EW70, n-Butanol fraction and ethyl acetate fraction, respectively. The results of the study suggest MF as a beneficial natural medicine.
{"title":"Antioxidant and antidiabetic potential of Myrothamnus flabellifolius found in Botswana","authors":"T. Kwape, R. Majinda, P. Chaturvedi","doi":"10.1080/23312025.2016.1275403","DOIUrl":"https://doi.org/10.1080/23312025.2016.1275403","url":null,"abstract":"Abstract The total phenol content, antioxidant and antidiabetic activity of Myrothamnus flabellifolius (MF) were investigated. The study revealed 70% ethanol/water (EW70), 70% methanol/water (MW70) and methanol (Me) extracts to contain the highest phenol content, 350 mg GAE/g, 300 mg GAE/g and 250 mg GAE/g, respectively (GAE/g—Gallic Acid Equivalent per gram). MW70 exhibited the highest radical [diphenyl picryl hydrazine (DPPH) and 2, 2-Azobis-3-ethyl benzthiazoline-6-sulphonic acid (ABTS+)] scavenging power. About 70% ethanol/water exhibited the highest reducing power, hydrogen peroxide and nitric oxide scavenging activities. The results of antidiabetic activity showed EW70 to possess the highest α-amylase inhibitory effect. The α-glucosidase activity was inhibited by EW70, n-Butanol fraction and ethyl acetate fraction, respectively. The results of the study suggest MF as a beneficial natural medicine.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1275403","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1225877
Kar-Yee Tye, S. Gan, S. Lim, S. Tan, Chen-Ai Chen, S. Phang
Abstract Marine natural products have been increasingly found to be a promising source of drug candidates for fighting human diseases. The present study was carried out to assess the antimicrobial properties of a brown alga, Turbinaria ornata. Hexane, dichloromethane, methanol, and water extracts were tested against 23 micro-organisms including Gram-positive and negative bacteria, yeasts, and fungi. The disk diffusion method was employed followed by modified resazurin microtitre assay (REMA). The results obtained from modified REMA using both methods of colorimetric and fluorometric were compared. The best antimicrobial activity was recorded in dichloromethane extract for disk diffusion. Further, modified REMA showed inhibition in Bacillus cereus, Bacillus subtilis, Staphylococcus aureus ATCC 25923, S. aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Enterococcus faecalis, Pseudomonas aeruginosa ATCC 27853, Candida parapsilosis ATCC 22019, Candida guilliermondii ATCC 6260, and Saccharomyces cerevisiae. Both methods of modified REMA were substantially in agreement with each other based on Cohen’s kappa statistical analysis (κ value = 0.712; p < 0.0005). Our findings suggested that T. ornata dichloromethane extract has the potential to be used as a source of antimicrobial compounds.
{"title":"Comparison of visual observation and emission intensity of resazurin for antimicrobial properties of hexane, dichloromethane, methanol and water extracts from a brown alga, Turbinaria ornata","authors":"Kar-Yee Tye, S. Gan, S. Lim, S. Tan, Chen-Ai Chen, S. Phang","doi":"10.1080/23312025.2016.1225877","DOIUrl":"https://doi.org/10.1080/23312025.2016.1225877","url":null,"abstract":"Abstract Marine natural products have been increasingly found to be a promising source of drug candidates for fighting human diseases. The present study was carried out to assess the antimicrobial properties of a brown alga, Turbinaria ornata. Hexane, dichloromethane, methanol, and water extracts were tested against 23 micro-organisms including Gram-positive and negative bacteria, yeasts, and fungi. The disk diffusion method was employed followed by modified resazurin microtitre assay (REMA). The results obtained from modified REMA using both methods of colorimetric and fluorometric were compared. The best antimicrobial activity was recorded in dichloromethane extract for disk diffusion. Further, modified REMA showed inhibition in Bacillus cereus, Bacillus subtilis, Staphylococcus aureus ATCC 25923, S. aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Enterococcus faecalis, Pseudomonas aeruginosa ATCC 27853, Candida parapsilosis ATCC 22019, Candida guilliermondii ATCC 6260, and Saccharomyces cerevisiae. Both methods of modified REMA were substantially in agreement with each other based on Cohen’s kappa statistical analysis (κ value = 0.712; p < 0.0005). Our findings suggested that T. ornata dichloromethane extract has the potential to be used as a source of antimicrobial compounds.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1225877","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1272166
S. Roy, B. Ukil, L. M. Lyndem
Abstract Senna alata has attracted the attention of many researchers due to its numerous medicinal properties. This study aims to test the acute and sub-acute toxicity of its leaf extracts in Swiss albino mice. Studies were carried out with a fixed dose of 1,000, 2,000, and 3,000 mg/kg body weight through oral administration daily. Signs of toxicity in terms of behavior and mortality were noted after every two hours till 24 h of administration for acute toxicity and further administration of extracts till 15 days to analyze the physical, biochemical, hematological parameters, and histopathological studies in liver, kidney, and spleen for sub-acute study. The highest dose administered did not produce mortality or changes in the general behavior of the test animals. All parameters were unaltered throughout the study. The present study revealed no obvious toxicity in mice treated with S. alata. These results indicate the safety of the oral administration of leaf extract.
{"title":"Acute and sub-acute toxicity studies on the effect of Senna alata in Swiss Albino mice","authors":"S. Roy, B. Ukil, L. M. Lyndem","doi":"10.1080/23312025.2016.1272166","DOIUrl":"https://doi.org/10.1080/23312025.2016.1272166","url":null,"abstract":"Abstract Senna alata has attracted the attention of many researchers due to its numerous medicinal properties. This study aims to test the acute and sub-acute toxicity of its leaf extracts in Swiss albino mice. Studies were carried out with a fixed dose of 1,000, 2,000, and 3,000 mg/kg body weight through oral administration daily. Signs of toxicity in terms of behavior and mortality were noted after every two hours till 24 h of administration for acute toxicity and further administration of extracts till 15 days to analyze the physical, biochemical, hematological parameters, and histopathological studies in liver, kidney, and spleen for sub-acute study. The highest dose administered did not produce mortality or changes in the general behavior of the test animals. All parameters were unaltered throughout the study. The present study revealed no obvious toxicity in mice treated with S. alata. These results indicate the safety of the oral administration of leaf extract.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1272166","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-31DOI: 10.1080/23312025.2016.1261577
Y. Wang, T. Engelmann, Y.F. Xu, W. Schwarz, T. Hsu
Abstract The GABAergic and glutamatergic systems play key roles in controlling activity of the central nervous system. Important membrane proteins in the mammalian central nervous system transporting extracellular GABA and glutamate are the GABA transporter GAT1 and the glutamate transporter EAAC1. We investigated the effect of catechins of green tea (Camellia sinensis) on the activity of GAT1 and EAAC1 by detecting the respective electrogenic transporter-mediated current under voltage clamp. Epigallocatechin-3-gallate inhibited GAT1-mediated current to 50% at about 100 μM. The EAAC1-mediated current could be stimulated up to 80% by (-)-epicatechin; 50% of maximum stimulation was achieved by about 5 μM. Inhibition of GAT1 and stimulation of EAAC1 will counteract hyperexcitability.
{"title":"Catechins from green tea modulate neurotransmitter transporter activity in Xenopus oocytes","authors":"Y. Wang, T. Engelmann, Y.F. Xu, W. Schwarz, T. Hsu","doi":"10.1080/23312025.2016.1261577","DOIUrl":"https://doi.org/10.1080/23312025.2016.1261577","url":null,"abstract":"Abstract The GABAergic and glutamatergic systems play key roles in controlling activity of the central nervous system. Important membrane proteins in the mammalian central nervous system transporting extracellular GABA and glutamate are the GABA transporter GAT1 and the glutamate transporter EAAC1. We investigated the effect of catechins of green tea (Camellia sinensis) on the activity of GAT1 and EAAC1 by detecting the respective electrogenic transporter-mediated current under voltage clamp. Epigallocatechin-3-gallate inhibited GAT1-mediated current to 50% at about 100 μM. The EAAC1-mediated current could be stimulated up to 80% by (-)-epicatechin; 50% of maximum stimulation was achieved by about 5 μM. Inhibition of GAT1 and stimulation of EAAC1 will counteract hyperexcitability.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1261577","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-17DOI: 10.1080/23312025.2016.1262489
A. A. Siddiqui, Peerzada Shariq Shaheen Khaki, A. Sohail, Tarique Sarwar, B. Bano
Abstract It is well known that fruit nuts contain wide variety of flavonoids and various proteins, consumption of which has been associated with the reduced risk of chronic diseases. Cystatins, a family of cysteine proteinase inhibitors, ubiquitously present in all cells serve various important and critical physiological functions. In this study a phytocystatin with molecular mass of 63.4 kDa was purified to homogeneity by a three-step process including ammonium sulfate fractionation (50–70%), acetone precipitation, and gel filtration chromatography on Sephacryl S100-HR column. The purified inhibitor migrated as single band under native and SDS-PAGE. The Ki values for purified inhibitor with papain, ficin, and bromelain were found to be 45.45, 83.33, and 90.9 nM, respectively, suggesting higher affinity of the inhibitor for papain as compared to ficin and bromelain. Phytocystatin was stable in broad pH and temperature range. Purified cystatin appeared to be antigenic as observed in western blot analysis. ITC assay data show a binding stoichiometry of 0.870 ± 0.03 sites for cystatin and papain interaction which indicated that cystatin is surrounded by nearly one papain molecule. FTIR, UV, and fluorescence studies showed significant conformational changes on cystatin–papain complex formation. Purified cystatin was found to possess 36.8% α-helical content as observed by CD spectroscopy.
{"title":"Isolation and purification of phytocystatin from almond: Biochemical, biophysical, and immunological characterization","authors":"A. A. Siddiqui, Peerzada Shariq Shaheen Khaki, A. Sohail, Tarique Sarwar, B. Bano","doi":"10.1080/23312025.2016.1262489","DOIUrl":"https://doi.org/10.1080/23312025.2016.1262489","url":null,"abstract":"Abstract It is well known that fruit nuts contain wide variety of flavonoids and various proteins, consumption of which has been associated with the reduced risk of chronic diseases. Cystatins, a family of cysteine proteinase inhibitors, ubiquitously present in all cells serve various important and critical physiological functions. In this study a phytocystatin with molecular mass of 63.4 kDa was purified to homogeneity by a three-step process including ammonium sulfate fractionation (50–70%), acetone precipitation, and gel filtration chromatography on Sephacryl S100-HR column. The purified inhibitor migrated as single band under native and SDS-PAGE. The Ki values for purified inhibitor with papain, ficin, and bromelain were found to be 45.45, 83.33, and 90.9 nM, respectively, suggesting higher affinity of the inhibitor for papain as compared to ficin and bromelain. Phytocystatin was stable in broad pH and temperature range. Purified cystatin appeared to be antigenic as observed in western blot analysis. ITC assay data show a binding stoichiometry of 0.870 ± 0.03 sites for cystatin and papain interaction which indicated that cystatin is surrounded by nearly one papain molecule. FTIR, UV, and fluorescence studies showed significant conformational changes on cystatin–papain complex formation. Purified cystatin was found to possess 36.8% α-helical content as observed by CD spectroscopy.","PeriodicalId":10412,"journal":{"name":"Cogent Biology","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/23312025.2016.1262489","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60091030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-14DOI: 10.1080/23312025.2016.1264691
M. E. Connor, T. R. King
Abstract Background: A combination of genetic fine-mapping and complementation testing was used previously to assign the juvenile alopecia mutation (abbreviated jal) to the GATA binding protein 3 (Gata3) gene on Chromosome 2 in mice. However, sequence analysis of Gata3 exons (including coding and noncoding regions) revealed no differences between wild type C3H/HeJ and co-isogenic C3H/HeJ-jal/J mutant mice. Results: Using a PCR-based scanning method, here we have tested the hypothesis that jal might result from insertion of a transposable element in or near the Gata3 gene. We show that the jal mutation is specifically associated with an intracisternal A particle (IAP) element of the I∆1 subtype that has transposed to Intron 3–4 in the Gata3 gene, and use the same panel of recombinants used previously to fine-map jal to show that this IAP element and jal are located within the same small genetic interval. Conclusion: Transposition of an IAP element of the I∆1 subtype into Intron 3–4 of the mutant Gata3jal allele is the likely cause of the juvenile alopecia phenotype in mutant mice.
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