Characterization of P-type ATPase 3 in Plasmodium falciparum
详细信息 查看全文
- 作者：Rozmajzl ; Patrick J. ; Kimura ; Masatsugu ; Woodrow ; Charles J. ; Krishna ; Sanjeev ; Meade ; John C.
- 关键词：P-type ATPase ; Plasmodium falcijparum ; Malaria ; PCR ; polymerase chain reaction ; UTR ; untranslated region ; nt ; nucleotide base ; RACE ; rapid amplification of cDNA ends ; TC-PCR ; tandem competitive polymerase chain reaction amplification ; aa ; amino acid
- 刊名：Molecular and Biochemical Parasitology
- 出版年：2001
- 出版时间：September, 2001
- 年：2001
- 卷：116
- 期：2
- 页码：117-126
- 全文大小：383 K
文摘
We report the nucleotide sequence, derived amino acid sequence and expression profile of P-type ATPase 3 (PfATPase3) from Plasmodium falciparum. An open reading frame of 7362 nucleotides, interrupted by a single intron of 168 nt, encoded a protein product of 2394 amino acids with a predicted MW of 282 791 Da. Hydropathy analysis of PfATPase3 revealed six amino-terminal and six carboxyl-terminal membrane spanning regions (M1–12) flanking a large hydrophilic domain with a smaller hydrophilic loop between M4 and M5. Based on a phylogenetic comparison of conserved domains present in P-type ATPases from other organisms, PfATPase3 resembled a Type-V ATPase for which the transport affinity is unknown. The PfATPase3 topology was interrupted by four regions, termed ‘inserts’, unique to malarial P-type ATPases, which were high in asparagine residues and charged amino acids (inserts I1–I4). Inserts I1 and I3 also contained repeated amino acid motifs. The number and composition of repeated amino acid motifs in insert I3 were variable in seven P. falciparum strains tested. PfATPase3 was 80.2 % similar to the non-insert portions of P. yoelii ATPase3, although their inserts differed in length and composition. PfATPase3 mRNA was most abundant relative to β-tubulin during the latter half of the erythrocytic cycle and was also present in gametocytes. Using affinity-purified antibody to a 14 amino acid PfATPase3 epitope, a 260 kDa protein was detected by Western analysis. Based on immunofluorescence, the PfATPase3 protein was located intracellularly in gametocytes and, to a lesser extent, in late erythrocytic stages.