Molecular and antigenic characterization of the Leishmania (Viannia) panamensis kinetoplastid membrane protein-11.
The kinetoplastid membrane protein 11 (KMP-11) has been recently described in Leishmania (Leishmania) donovani as a major component of the promastigote membrane. Two oligonucleotide primers were synthesized to PCR-amplify the entire encoding region of New World Leishmania species. The Leishmania (Viannia) panamensis amplification product was clone, sequenced and the putative amino acid sequence determined. A remarkably high degree of sequence homology was observed with the corresponding molecule of L. (L) donovani and L. (L) infantum (97% and 96%, respectively). Southern blot analysis showed that the KMP-11 locus is conformed by three copies of the gene. the L. (V) panamensis ORF was subsequently clone in a high expression vector and the recombinant protein was induced and purified from Escherichia coli cultures. Immunoblot analysis showed that 80%, 77% and 100% sera from cutaneous, mucocutaneous and visceral leishmaniasis patients, respectively, recognized the recombinant KMP-11 protein. In a similar assay, 86% of asymptomatic Leishmania-infected individuals showed IgG antibodies against the rKMP-11. We proposed that KMP-11 could be used as a serologic marker for infection and disease caused by Leishmania in America.
Ramírez
J R
JR
Programa de Estudio y Control de Enfermedades Tropicales, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia.
Berberich
C
C
Jaramillo
A
A
Alonso
C
C
Vélez
I V
IV
D000818:Animals; D000911:Antibodies, Monoclonal; D015870:Gene Expression; D016678:Genome; D006801:Humans; D018334:Leishmania guyanensis / Q000737:chemistry*; D007896:Leishmaniasis; D008565:Membrane Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D011994:Recombinant Proteins