Comparative protein profiling identifies elongation factor-1beta and tryparedoxin peroxidase as factors associated with metastasis in Leishmania guyanensis.
Parasites of the Leishmania Viannia subgenus are major causative agents of mucocutaneous leishmaniasis (MCL), a disease characterised by parasite dissemination (metastasis) from the original cutaneous lesion to form debilitating secondary lesions in the nasopharyngeal mucosa. We employed a protein profiling approach to identify potential metastasis factors in laboratory clones of L. (V.) guyanensis with stable phenotypes ranging from highly metastatic (M+) through infrequently metastatic (M+/M-) to non-metastatic (M-). Comparison of the soluble proteomes of promastigotes by two-dimensional electrophoresis revealed two abundant protein spots specifically associated with M+ and M+/M- clones (Met2 and Met3) and two others exclusively expressed in M- parasites (Met1 and Met4). The association between clinical disease phenotype and differential expression of Met1-Met4 was less clear in L. Viannia strains from mucosal (M+) or cutaneous (M-) lesions of patients. Identification of Met1-Met4 by biological mass spectrometry (LC-ES-MS/MS) and bioinformatics revealed that M+ and M- clones express distinct acidic and neutral isoforms of both elongation factor-1 subunit beta (EF-1beta) and cytosolic tryparedoxin peroxidase (TXNPx). This interchange of isoforms may relate to the mechanisms by which the activities of EF-1beta and TXNPx are modulated, and/or differential post-translational modification of the gene product(s). The multiple metabolic functions of EF-1 and TXNPx support the plausibility of their participation in parasite survival and persistence and thereby, metastatic disease. Both polypeptides are active in resistance to chemical and oxidant stress, providing a basis for further elucidation of the importance of antioxidant defence in the pathogenesis underlying MCL.
Walker
John
J
Centro Internacional de Entrenamiento e Investigaciones Medicas (CIDEIM), Avenida 1 Norte No. 3-03, Cali, Colombia. john_walker@cideim.org.co
Acestor
Nathalie
N
Gongora
Rafael
R
Quadroni
Manfredo
M
Segura
Iris
I
Fasel
Nicolas
N
Saravia
Nancy G
NG
D000595:Amino Acid Sequence; D000818:Animals; D019295:Computational Biology; D016054:DNA, Protozoan; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D015151:Immunoblotting; D018334:Leishmania guyanensis / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1 / Q000032:analysis*; D010544:Peroxidases / Q000032:analysis*; D020033:Protein Isoforms / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D016054:DNA, Protozoan; D020648:Peptide Elongation Factor 1; D020033:Protein Isoforms; D020543:Proteome; D015800:Protozoan Proteins; D010544:Peroxidases; C110959:tryparedoxin peroxidase