The malaria parasite Plasmodium falciparum has only one pyruvate dehydrogenase complex, which is located in the apicoplast

被引:131
作者
Foth, BJ
Stimmler, LM
Handman, E
Crabb, BS
Hodder, AN
McFadden, GI [1 ]
机构
[1] Univ Melbourne, Sch Bot, Plant Cell Biol Res Ctr, Parkville, Vic 3010, Australia
[2] Royal Melbourne Hosp, Walter & Eliza Hall Inst Med Res, Dept Infect & Immun, Parkville, Vic 3050, Australia
关键词
D O I
10.1111/j.1365-2958.2004.04407.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The relict plastid (apicoplast) of apicomplexan parasites synthesizes fatty acids and is a promising drug target. In plant plastids, a pyruvate dehydrogenase complex (PDH) converts pyruvate into acetyl-CoA, the major fatty acid precursor, whereas a second, distinct PDH fuels the tricarboxylic acid cycle in the mitochondria. In contrast, the presence of genes encoding PDH and related enzyme complexes in the genomes of five Plasmodium species and of Toxoplasma gondii indicate that these parasites contain only one single PDH. PDH complexes are comprised of four subunits (E1alpha, E1beta, E2, E3), and we confirmed four genes encoding a complete PDH in Plasmodium falciparum through sequencing of cDNA clones. In apicomplexan parasites, many nuclear-encoded proteins are targeted to the apicoplast courtesy of two-part N-terminal leader sequences, and the presence of such N-terminal sequences on all four PDH subunits as well as phylogenetic analyses strongly suggest that the P. falciparum PDH is located in the apicoplast. Fusion of the two-part leader sequences from the E1alpha and E2 genes to green fluorescent protein experimentally confirmed apicoplast targeting. Western blot analysis provided evidence for the expression of the E1alpha and E1beta PDH subunits in blood-stage malaria parasites. The recombinantly expressed catalytic domain of the PDH subunit E2 showed high enzymatic activity in vitro indicating that pyruvate is converted to acetyl-CoA in the apicoplast, possibly for use in fatty acid biosynthesis.
引用
收藏
页码:39 / 53
页数:15
相关论文
共 64 条
[1]   Crystal structure of human branched-chain α-ketoacid dehydrogenase and the molecular basis of multienzyme complex deficiency in maple syrup urine disease [J].
AEvarsson, A ;
Chuang, JL ;
Wynn, RM ;
Turley, S ;
Chuang, DT ;
Hol, WGJ .
STRUCTURE, 2000, 8 (03) :277-291
[2]  
AEvarsson A, 1999, NAT STRUCT BIOL, V6, P785
[3]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[4]   Overcoming codon bias:: A method for high-level overexpression of Plasmodium and other AT-rich parasite genes in Escherichia coli [J].
Baca, AM ;
Hol, WGJ .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 2000, 30 (02) :113-118
[5]   Understanding in vivo carbon precursor supply for fatty acid synthesis in leaf tissue [J].
Bao, XM ;
Focke, M ;
Pollard, M ;
Ohlrogge, J .
PLANT JOURNAL, 2000, 22 (01) :39-50
[6]   Methionine regeneration and aspartate aminotransferase in parasitic protozoa [J].
Berger, LC ;
Wilson, J ;
Wood, P ;
Berger, BJ .
JOURNAL OF BACTERIOLOGY, 2001, 183 (15) :4421-4434
[7]   The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum [J].
Bozdech, Z ;
Llinás, M ;
Pulliam, BL ;
Wong, ED ;
Zhu, JC ;
DeRisi, JL .
PLOS BIOLOGY, 2003, 1 (01) :85-100
[8]   Genome sequence and comparative analysis of the model rodent malaria parasite Plasmodium yoelii yoelii [J].
Carlton, JM ;
Angiuoli, SV ;
Suh, BB ;
Kooij, TW ;
Pertea, M ;
Silva, JC ;
Ermolaeva, MD ;
Allen, JE ;
Selengut, JD ;
Koo, HL ;
Peterson, JD ;
Pop, M ;
Kosack, DS ;
Shumway, MF ;
Bidwell, SL ;
Shallom, SJ ;
van Aken, SE ;
Riedmuller, SB ;
Feldblyum, TV ;
Cho, JK ;
Quackenbush, J ;
Sedegah, M ;
Shoaibi, A ;
Cummings, LM ;
Florens, L ;
Yates, JR ;
Raine, JD ;
Sinden, RE ;
Harris, MA ;
Cunningham, DA ;
Preiser, PR ;
Bergman, LW ;
Vaidya, AB ;
Van Lin, LH ;
Janse, CJ ;
Waters, AP ;
Smith, HO ;
White, OR ;
Salzberg, SL ;
Venter, JC ;
Fraser, CM ;
Hoffman, SL ;
Gardner, MJ ;
Carucci, DJ .
NATURE, 2002, 419 (6906) :512-519
[9]  
CASPERS P, 1994, CELL MOL BIOL, V40, P635
[10]   Structural basis for flip-flop action of thiamin pyrophosphate-dependent enzymes revealed by human pyruvate dehydrogenase [J].
Ciszak, EM ;
Korotchkina, LG ;
Dominiak, PM ;
Sidhu, S ;
Patel, MS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (23) :21240-21246