Triosephosphate isomerase （TPI） catalyzes the interconversion of glyceraldehyde-3-phosphate to dihydroxyacetone phosphate. Photosynthetic organisms generally contain two isoforms of TPI located in both cytoplasm and...Triosephosphate isomerase （TPI） catalyzes the interconversion of glyceraldehyde-3-phosphate to dihydroxyacetone phosphate. Photosynthetic organisms generally contain two isoforms of TPI located in both cytoplasm and chloroplasts. While the cytoplasmic TPI is involved in the glycolysis, the chloroplastic isoform participates in the Calvin-Benson cycle, a key photosynthetic process responsible for carbon fixation. Compared with its cytoplasmic counterpart, the functional features of chloroplastic TPI have been poorly investigated and its three-dimensional structure has not been solved. Recently, several studies proposed TPI as a potential target of different redox modifications including dithiol/disulfide interchanges, glutathionylation, and nitrosylation. However, neither the effects on protein activity nor the molecular mechanisms underlying these redox modifications have been investigated. Here, we have produced recombinantly and purified TPI from the unicellular green alga Chlamydomonas reinhardtii （Cr）. The biochemical properties of the enzyme were delineated and its crystallographic structure was determined at a resolution of 1.1 A. CrTPI is a homodimer with subunits containing the typical （β/α）8-barrel fold. Although no evidence for TRX regulation was obtained, CrTPI was found to undergo glutathionylation by oxidized glutathione and trans-nitrosylation by nitrosoglutathione, confirming its sensitivity to multiple redox modifications.展开更多
Background:Giardia duodenalis is a species complex consisting of multiple genetically distinct assemblages.The species imposes a major public health crisis on developing countries.However,the molecular diversity,trans...Background:Giardia duodenalis is a species complex consisting of multiple genetically distinct assemblages.The species imposes a major public health crisis on developing countries.However,the molecular diversity,transmission dynamics and risk factors of the species in these countries are indeterminate.This study was conducted to determine the molecular epidemiology of G.duodenalis infection in asymptomatic individuals in Southern Ethiopia.Methods:From March to June 2014,fresh stool samples were collected from 590 randomly selected individuals.Socio-demographic data were gathered using a pre-tested structured questionnaire.The genotyping was done using triosephosphate isomerase gene-based nested polymerase chain reaction and DNA sequencing.The genetic identity and relatedness of isolates were determined using the basic local alignment search tool and phylogenetic analysis.Risk factors associated with G.duodenalis infection were analysed using binary and multinomial logistic regression models.Results:The results showed that 18.1%(92/509)of the study subjects were infected by G.duodenalis.Among the isolates,35.9%(33/92)and 21.7%(20/92)were sub-typed into assemblages A and B,respectively,whereas 42.4%(39/92)showed mixed infections of A and B.Most of the assemblage A isolates(94%,31/33)were 100%identical to sequences registered in GenBank,of which the majority belonged to sub-assemblage AII.However,the high genetic variability and frequency of double peaks made sub-genotyping of assemblage B more problematic and only 20%(4/20)of the isolates matched 100%with the sequences.The risk factors of age(P=0.032)and type of drinking water source(P=0.003)both showed a significant association with the occurrence G.duodenalis infection.Conclusions:This study established the endemicity of G.duodenalis in Southern Ethiopia.Infection with assemblage A was more frequent than with assemblage B,and the rate of infection was higher in children and in municipal/tap and open spring water consumers than the other groups.Sub-typing of assemblage B and determining the origin of double peaks were challenging.The present study confirms the need for further inclusive studies to be conducted focusing on sub-types of assemblage B and the origin of heterogeneity.展开更多
The parasite Plasmodium falciparum is responsible for the major world scourge malaria, a disease that affects 3.3 billion people worldwide. The development of new drugs is critical because of the diminished effectiven...The parasite Plasmodium falciparum is responsible for the major world scourge malaria, a disease that affects 3.3 billion people worldwide. The development of new drugs is critical because of the diminished effectiveness of current antimalarial agents mainly due to parasitic resistance, side effects and cost. Molecular docking was used to explore structural motifs responsible for the interactions between triose phosphate isomerase (TPI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and aldolase (ALD) from human and Plasmodium cells with 8 novel sufonylamide derivatives. All the ligands modeled, interact with all three enzymes in the micromolar range. The top ligand (sulfaE) shows a 70-fold increase in selective binding to pfTPI compared to hTPI (dissociation constant-KI of 7.83 μM and 0.177 μM for hTPI and pfTPI respectively), on par with antimalarial drug chloroquine.ALD and GAPDH form complexes with similar binding sites, comprising amino acids of similar chemical properties and polarities. Human TPI and pfTPI bind sulfonamide derivatives using two distinct binding sites and residues. Key residues at the dimer interface of pfTPI (VAL44, SER45, TYR48, GLN64, ASN65, VAL78) form a tight pocket with favorable polar contacts. The affinity with TPI is the most specific, stable, and selective suggesting pfTPI is a candidate for development of antimalarial drugs.展开更多
为了进一步认识褐虾中的一种过敏原——磷酸丙糖异构酶过敏原(Cra c 8)的结构与功能关系,理解食物过敏原之间产生交叉反应的分子基础,采用生物信息学方法分析Cra c 8蛋白的理化性质和各级结构,探讨其与食物中该蛋白质在分子水平上的异...为了进一步认识褐虾中的一种过敏原——磷酸丙糖异构酶过敏原(Cra c 8)的结构与功能关系,理解食物过敏原之间产生交叉反应的分子基础,采用生物信息学方法分析Cra c 8蛋白的理化性质和各级结构,探讨其与食物中该蛋白质在分子水平上的异同。结果表明:褐虾Cra c 8蛋白是由249个氨基酸组成的酸性蛋白质,与螯虾、对虾、三文鱼、小麦、蟑螂中的磷酸丙糖异构酶蛋白具有较高的同源性,亲疏水性区域相似;二级结构预测结果显示其以α-螺旋和不规则折叠为主,均没有β-折叠及β-转角区域;利用同源建模的方式成功的构建褐虾Cra c 8蛋白的空间结构。展开更多
文摘Triosephosphate isomerase （TPI） catalyzes the interconversion of glyceraldehyde-3-phosphate to dihydroxyacetone phosphate. Photosynthetic organisms generally contain two isoforms of TPI located in both cytoplasm and chloroplasts. While the cytoplasmic TPI is involved in the glycolysis, the chloroplastic isoform participates in the Calvin-Benson cycle, a key photosynthetic process responsible for carbon fixation. Compared with its cytoplasmic counterpart, the functional features of chloroplastic TPI have been poorly investigated and its three-dimensional structure has not been solved. Recently, several studies proposed TPI as a potential target of different redox modifications including dithiol/disulfide interchanges, glutathionylation, and nitrosylation. However, neither the effects on protein activity nor the molecular mechanisms underlying these redox modifications have been investigated. Here, we have produced recombinantly and purified TPI from the unicellular green alga Chlamydomonas reinhardtii （Cr）. The biochemical properties of the enzyme were delineated and its crystallographic structure was determined at a resolution of 1.1 A. CrTPI is a homodimer with subunits containing the typical （β/α）8-barrel fold. Although no evidence for TRX regulation was obtained, CrTPI was found to undergo glutathionylation by oxidized glutathione and trans-nitrosylation by nitrosoglutathione, confirming its sensitivity to multiple redox modifications.
基金The proposal development,study design and data collection of this research was supported and granted by Arba Minch University and Jimma University,Ethiopia.Erasmus Mundus Action 2 program(CARIBU)financially supported laboratory works of this study to be carried out at the Cell and Genetics(CEGE)Laboratory of Vrije Universiteit Brussel(VUB),Belgium.VUB granted and permitted the laboratory activities to be successfully carried out and completed。
文摘Background:Giardia duodenalis is a species complex consisting of multiple genetically distinct assemblages.The species imposes a major public health crisis on developing countries.However,the molecular diversity,transmission dynamics and risk factors of the species in these countries are indeterminate.This study was conducted to determine the molecular epidemiology of G.duodenalis infection in asymptomatic individuals in Southern Ethiopia.Methods:From March to June 2014,fresh stool samples were collected from 590 randomly selected individuals.Socio-demographic data were gathered using a pre-tested structured questionnaire.The genotyping was done using triosephosphate isomerase gene-based nested polymerase chain reaction and DNA sequencing.The genetic identity and relatedness of isolates were determined using the basic local alignment search tool and phylogenetic analysis.Risk factors associated with G.duodenalis infection were analysed using binary and multinomial logistic regression models.Results:The results showed that 18.1%(92/509)of the study subjects were infected by G.duodenalis.Among the isolates,35.9%(33/92)and 21.7%(20/92)were sub-typed into assemblages A and B,respectively,whereas 42.4%(39/92)showed mixed infections of A and B.Most of the assemblage A isolates(94%,31/33)were 100%identical to sequences registered in GenBank,of which the majority belonged to sub-assemblage AII.However,the high genetic variability and frequency of double peaks made sub-genotyping of assemblage B more problematic and only 20%(4/20)of the isolates matched 100%with the sequences.The risk factors of age(P=0.032)and type of drinking water source(P=0.003)both showed a significant association with the occurrence G.duodenalis infection.Conclusions:This study established the endemicity of G.duodenalis in Southern Ethiopia.Infection with assemblage A was more frequent than with assemblage B,and the rate of infection was higher in children and in municipal/tap and open spring water consumers than the other groups.Sub-typing of assemblage B and determining the origin of double peaks were challenging.The present study confirms the need for further inclusive studies to be conducted focusing on sub-types of assemblage B and the origin of heterogeneity.
文摘The parasite Plasmodium falciparum is responsible for the major world scourge malaria, a disease that affects 3.3 billion people worldwide. The development of new drugs is critical because of the diminished effectiveness of current antimalarial agents mainly due to parasitic resistance, side effects and cost. Molecular docking was used to explore structural motifs responsible for the interactions between triose phosphate isomerase (TPI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and aldolase (ALD) from human and Plasmodium cells with 8 novel sufonylamide derivatives. All the ligands modeled, interact with all three enzymes in the micromolar range. The top ligand (sulfaE) shows a 70-fold increase in selective binding to pfTPI compared to hTPI (dissociation constant-KI of 7.83 μM and 0.177 μM for hTPI and pfTPI respectively), on par with antimalarial drug chloroquine.ALD and GAPDH form complexes with similar binding sites, comprising amino acids of similar chemical properties and polarities. Human TPI and pfTPI bind sulfonamide derivatives using two distinct binding sites and residues. Key residues at the dimer interface of pfTPI (VAL44, SER45, TYR48, GLN64, ASN65, VAL78) form a tight pocket with favorable polar contacts. The affinity with TPI is the most specific, stable, and selective suggesting pfTPI is a candidate for development of antimalarial drugs.
文摘为了进一步认识褐虾中的一种过敏原——磷酸丙糖异构酶过敏原(Cra c 8)的结构与功能关系,理解食物过敏原之间产生交叉反应的分子基础,采用生物信息学方法分析Cra c 8蛋白的理化性质和各级结构,探讨其与食物中该蛋白质在分子水平上的异同。结果表明:褐虾Cra c 8蛋白是由249个氨基酸组成的酸性蛋白质,与螯虾、对虾、三文鱼、小麦、蟑螂中的磷酸丙糖异构酶蛋白具有较高的同源性,亲疏水性区域相似;二级结构预测结果显示其以α-螺旋和不规则折叠为主,均没有β-折叠及β-转角区域;利用同源建模的方式成功的构建褐虾Cra c 8蛋白的空间结构。
