Analysis of NLS and rRNA binding motifs in the L25 ribosomal protein from Leishmania (viannia) braziliensis: investigation of its diagnostic capabilities.
A cDNA clone codifying ribosomal protein L25 was isolated from a Leishmania braziliensis cDNA gene library. The alignment of the amino acid sequence deduced from this gene with other proteins revealed that this protein is related to the L23/25 rihosomal protein family. This is so because this protein shows, in its C-terminal end, the rRNA binding domains characteristic of these proteins and at the N-terminal end the NLS sequence necessary for its entry into the nucleus. Southern blot analysis showed 2 copies of gene L25 per genome arranged in tandem position and pointing in the same direction. Northern blot analysis showed that this gene is transcribed in 2 mRNAs when parasite promastigotes are in the logarithmic phase. In order to analyse the antigenic properties of L. braziliensis RPL25, it was purified as a recombinant protein and ELISA-tested against cutaneous, mucocutaneous and Chagasic sera. The results indicate that the recombinant RPL25 from L. braziliensis presents a non-specific reaction that disqualifies it for the diagnosis of cutaneous leishmaniasis. In contrast, some of the synthetic peptides derived from its sequence may serve as promising tools for the diagnosis of this disease.
González
A C
AC
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Tenerife, Spain.
Martínez
E
E
Carmelo
E
E
Piñero
J E
JE
Alonso
V
V
Del Castillo
A
A
Valladares
B
B
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D014355:Chagas Disease / Q000276:immunology / Q000469:parasitology; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007892:Leishmania braziliensis / Q000235:genetics* / Q000378:metabolism; D016773:Leishmaniasis, Cutaneous / Q000276:immunology / Q000469:parasitology; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D000913:Antibodies, Protozoan; D016054:DNA, Protozoan; D019913:Nuclear Localization Signals; D012335:RNA, Ribosomal; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C031127:ribosomal protein L25