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Expression of a MAPEG GST from Leishmania tarentolae
Author(s) -
Osei-Owusu Nana Yaw,
Hearne Jennifer,
Currington Rashaila
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1112.5
Subject(s) - heme , leishmania , biology , biochemistry , gene , heme oxygenase , enzyme , genetics , chemistry , parasite hosting , world wide web , computer science
Leishmaniasis is a tropical disease that afflicts approximately 12 million of the world's population. It causative agent is the protozoa Leishmania, a member of the Trypanosomatidea family. Trypanosomes lack a complete heme biosynthetic pathway; therefore, they must acquire heme exogenously. Its mechanism of heme scavenging is unclear. One way of heme acquisition may be accomplished through the coordination of heme with GSH via MAPEG, members of the GST superfamily. We know that Macaca fascicularis microsomal prostaglandin E synthase type 2, a MAPEG GST, binds GSH which forms a coordination bond with heme. We hypothesized that there is an enzyme similar to mPGES‐ 2 which scavenges heme in Leishmania. We then identified the first GST member gene, resembling mPGES‐2, in Leishmania. Identification of the target gene in the L. major genome was accomplished using BLAST at GeneDB. Amplified homolog in L. tarentolae was ligated with the pCR8⁄GW⁄TOPO TA vector for recombination with pDEST14 to generate an expression vector. Since Leishmania lack a complete heme biosynthetic pathway, exploitation of its heme dependency is a key target for drug discovery