2ayv: Difference between revisions

Revision as of 17:32, 21 February 2008

Crystal structure of a putative ubiquitin-conjugating enzyme E2 from Toxoplasma gondii

OverviewOverview

Parasites from the protozoan phylum Apicomplexa are responsible for diseases, such as malaria, toxoplasmosis and cryptosporidiosis, all of which have significantly higher rates of mortality and morbidity in economically underdeveloped regions of the world. Advances in vaccine development and drug discovery are urgently needed to control these diseases and can be facilitated by production of purified recombinant proteins from Apicomplexan genomes and determination of their 3D structures. To date, both heterologous expression and crystallization of Apicomplexan proteins have seen only limited success. In an effort to explore the effectiveness of producing and crystallizing proteins on a genome-scale using a standardized methodology, over 400 distinct Plasmodium falciparum target genes were chosen representing different cellular classes, along with select orthologues from four other Plasmodium species as well as Cryptosporidium parvum and Toxoplasma gondii. From a total of 1008 genes from the seven genomes, 304 (30.2%) produced purified soluble proteins and 97 (9.6%) crystallized, culminating in 36 crystal structures. These results demonstrate that, contrary to previous findings, a standardized platform using Escherichia coli can be effective for genome-scale production and crystallography of Apicomplexan proteins. Predictably, orthologous proteins from different Apicomplexan genomes behaved differently in expression, purification and crystallization, although the overall success rates of Plasmodium orthologues do not differ significantly. Their differences were effectively exploited to elevate the overall productivity to levels comparable to the most successful ongoing structural genomics projects: 229 of the 468 target genes produced purified soluble protein from one or more organisms, with 80 and 32 of the purified targets, respectively, leading to crystals and ultimately structures from one or more orthologues.

About this StructureAbout this Structure

2AYV is a Protein complex structure of sequences from Toxoplasma gondii with as ligand. Active as Ubiquitin--protein ligase, with EC number 6.3.2.19 Full crystallographic information is available from OCA.

ReferenceReference

Genome-scale protein expression and structural biology of Plasmodium falciparum and related Apicomplexan organisms., Vedadi M, Lew J, Artz J, Amani M, Zhao Y, Dong A, Wasney GA, Gao M, Hills T, Brokx S, Qiu W, Sharma S, Diassiti A, Alam Z, Melone M, Mulichak A, Wernimont A, Bray J, Loppnau P, Plotnikova O, Newberry K, Sundararajan E, Houston S, Walker J, Tempel W, Bochkarev A, Kozieradzki I, Edwards A, Arrowsmith C, Roos D, Kain K, Hui R, Mol Biochem Parasitol. 2007 Jan;151(1):100-10. Epub 2006 Nov 13. PMID:17125854

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OCA

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 ==Overview==
-Parasites from the protozoan phylum Apicomplexa are responsible for, diseases, such as malaria, toxoplasmosis and cryptosporidiosis, all of, which have significantly higher rates of mortality and morbidity in, economically underdeveloped regions of the world. Advances in vaccine, development and drug discovery are urgently needed to control these, diseases and can be facilitated by production of purified recombinant, proteins from Apicomplexan genomes and determination of their 3D, structures. To date, both heterologous expression and crystallization of, Apicomplexan proteins have seen only limited success. In an effort to, explore the effectiveness of producing and crystallizing proteins on a, genome-scale using a standardized methodology, over 400 distinct, Plasmodium falciparum target genes were chosen representing different, cellular classes, along with select orthologues from four other Plasmodium, species as well as Cryptosporidium parvum and Toxoplasma gondii. From a, total of 1008 genes from the seven genomes, 304 (30.2%) produced purified, soluble proteins and 97 (9.6%) crystallized, culminating in 36 crystal, structures. These results demonstrate that, contrary to previous findings, a standardized platform using Escherichia coli can be effective for, genome-scale production and crystallography of Apicomplexan proteins., Predictably, orthologous proteins from different Apicomplexan genomes, behaved differently in expression, purification and crystallization, although the overall success rates of Plasmodium orthologues do not differ, significantly. Their differences were effectively exploited to elevate the, overall productivity to levels comparable to the most successful ongoing, structural genomics projects: 229 of the 468 target genes produced, purified soluble protein from one or more organisms, with 80 and 32 of the, purified targets, respectively, leading to crystals and ultimately, structures from one or more orthologues.
+Parasites from the protozoan phylum Apicomplexa are responsible for diseases, such as malaria, toxoplasmosis and cryptosporidiosis, all of which have significantly higher rates of mortality and morbidity in economically underdeveloped regions of the world. Advances in vaccine development and drug discovery are urgently needed to control these diseases and can be facilitated by production of purified recombinant proteins from Apicomplexan genomes and determination of their 3D structures. To date, both heterologous expression and crystallization of Apicomplexan proteins have seen only limited success. In an effort to explore the effectiveness of producing and crystallizing proteins on a genome-scale using a standardized methodology, over 400 distinct Plasmodium falciparum target genes were chosen representing different cellular classes, along with select orthologues from four other Plasmodium species as well as Cryptosporidium parvum and Toxoplasma gondii. From a total of 1008 genes from the seven genomes, 304 (30.2%) produced purified soluble proteins and 97 (9.6%) crystallized, culminating in 36 crystal structures. These results demonstrate that, contrary to previous findings, a standardized platform using Escherichia coli can be effective for genome-scale production and crystallography of Apicomplexan proteins. Predictably, orthologous proteins from different Apicomplexan genomes behaved differently in expression, purification and crystallization, although the overall success rates of Plasmodium orthologues do not differ significantly. Their differences were effectively exploited to elevate the overall productivity to levels comparable to the most successful ongoing structural genomics projects: 229 of the 468 target genes produced purified soluble protein from one or more organisms, with 80 and 32 of the purified targets, respectively, leading to crystals and ultimately structures from one or more orthologues.
 ==About this Structure==
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 [[Category: Lew, J.]]
 [[Category: Melone, M.]]
-[[Category: SGC, Structural.Genomics.Consortium.]]
+[[Category: SGC, Structural Genomics Consortium.]]
 [[Category: Sundstrom, M.]]
 [[Category: Tempel, W.]]
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 [[Category: ubiquitin-conjugating enzyme]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jan 29 18:12:51 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:32:25 2008''