title: Genome-scale protein expression and structural biology of Plasmodium falciparum and related Apicomplexan organisms.
abstract: 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.
authors.affiliation: Structural Genomics Consortium, U. of Toronto, 100 College St. Rm 522B, Toronto, Ont., Canada M5G 1L5.
authors.forename: Masoud
authors.initials: M
authors.lastname: Vedadi
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Lew
authors.forename: Jennifer
authors.initials: J
authors.lastname: Artz
authors.forename: Mehrnaz
authors.initials: M
authors.lastname: Amani
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Aiping
authors.initials: A
authors.lastname: Dong
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Wasney
authors.forename: Mian
authors.initials: M
authors.lastname: Gao
authors.forename: Tanya
authors.initials: T
authors.lastname: Hills
authors.forename: Stephen
authors.initials: S
authors.lastname: Brokx
authors.forename: Wei
authors.initials: W
authors.lastname: Qiu
authors.forename: Sujata
authors.initials: S
authors.lastname: Sharma
authors.forename: Angelina
authors.initials: A
authors.lastname: Diassiti
authors.forename: Zahoor
authors.initials: Z
authors.lastname: Alam
authors.forename: Michelle
authors.initials: M
authors.lastname: Melone
authors.forename: Anne
authors.initials: A
authors.lastname: Mulichak
authors.forename: Amy
authors.initials: A
authors.lastname: Wernimont
authors.forename: James
authors.initials: J
authors.lastname: Bray
authors.forename: Peter
authors.initials: P
authors.lastname: Loppnau
authors.forename: Olga
authors.initials: O
authors.lastname: Plotnikova
authors.forename: Kate
authors.initials: K
authors.lastname: Newberry
authors.forename: Emayavaram
authors.initials: E
authors.lastname: Sundararajan
authors.forename: Simon
authors.initials: S
authors.lastname: Houston
authors.forename: John
authors.initials: J
authors.lastname: Walker
authors.forename: Wolfram
authors.initials: W
authors.lastname: Tempel
authors.forename: Alexey
authors.initials: A
authors.lastname: Bochkarev
authors.forename: Ivona
authors.initials: I
authors.lastname: Kozieradzki
authors.forename: Aled
authors.initials: A
authors.lastname: Edwards
authors.forename: Cheryl
authors.initials: C
authors.lastname: Arrowsmith
authors.forename: David
authors.initials: D
authors.lastname: Roos
authors.forename: Kevin
authors.initials: K
authors.lastname: Kain
authors.forename: Raymond
authors.initials: R
authors.lastname: Hui
mesh_terms: D000818:Animals; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018503:Genome, Protozoan / Q000235:genetics*; D008958:Models, Molecular; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012995:Solubility
chemical_list: D015800:Protozoan Proteins