Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells

Martha Forero, Álvaro Puentes, Jimena Cortés, Fabio Castillo, Ricardo Vera, Luis E. Rodríguez, John Valbuena, Marisol Ocampo, Hernando Curtidor, Jaiver Rosas, Javier García, Gloria Barrera, Rosalba Alfonso, Manuel A. Patarroyo, Manuel E. Patarroyo

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Virulence and immunity are still poorly understood in Mycobacterium tuberculosis. The H37Rv M. tuberculosis laboratory strain genome has been completely sequenced, and this along with proteomic technology represent powerful tools contributing toward studying the biology of target cell interaction with a facultative bacillus and designing new strategies for controlling tuberculosis. Rv2004c is a putative M. tuberculosis protein that could have specific mycobacterial functions. This study has revealed that the encoding gene is present in all mycobacterium species belonging to the M. tuberculosis complex. Rv2004c gene transcription was observed in all of this complex's strains except Mycobacterium bovis and Mycobacterium microti. Rv2004c protein expression was confirmed by using antibodies able to recognize a 54-kDa molecule by immunoblotting, and its location was detected on the M. tuberculosis surface by transmission electron microscopy, suggesting that it is a mycobacterial surface protein. Binding assays led to recognizing high activity binding peptides (HABP); five HABPs specifically bound to U937 cells, and six specifically bound to A549 cells. HABP circular dichroism suggested that they had an α-helical structure. HABP-target cell interaction was determined to be specific and saturable; some of them also displayed greater affinity for A549 cells than U937 cells. The critical amino acids directly involved in their interaction with U937 cells were also determined. Two probable receptor molecules were found on U937 cells and five on A549 for the two HABPs analyzed. These observations have important biological significance for studying bacillus-target cell interactions and implications for developing strategies for controlling this disease. Copyright © 2005 The Protein Society.
Original languageEnglish (US)
Pages (from-to)2767-2780
Number of pages14
JournalProtein Science
DOIs
StatePublished - Nov 1 2005

Fingerprint

Macrophages
U937 Cells
Mycobacterium tuberculosis
Epithelial Cells
Bacilli
Peptides
Cell Communication
Genes
Mycobacterium
Proteins
Molecules
Gene encoding
Bacillus
Transcription
Assays
Membrane Proteins
Arvicolinae
Transmission electron microscopy
Amino Acids
Mycobacterium bovis

Cite this

Forero, Martha ; Puentes, Álvaro ; Cortés, Jimena ; Castillo, Fabio ; Vera, Ricardo ; Rodríguez, Luis E. ; Valbuena, John ; Ocampo, Marisol ; Curtidor, Hernando ; Rosas, Jaiver ; García, Javier ; Barrera, Gloria ; Alfonso, Rosalba ; Patarroyo, Manuel A. ; Patarroyo, Manuel E. / Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells. In: Protein Science. 2005 ; pp. 2767-2780.
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title = "Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells",
abstract = "Virulence and immunity are still poorly understood in Mycobacterium tuberculosis. The H37Rv M. tuberculosis laboratory strain genome has been completely sequenced, and this along with proteomic technology represent powerful tools contributing toward studying the biology of target cell interaction with a facultative bacillus and designing new strategies for controlling tuberculosis. Rv2004c is a putative M. tuberculosis protein that could have specific mycobacterial functions. This study has revealed that the encoding gene is present in all mycobacterium species belonging to the M. tuberculosis complex. Rv2004c gene transcription was observed in all of this complex's strains except Mycobacterium bovis and Mycobacterium microti. Rv2004c protein expression was confirmed by using antibodies able to recognize a 54-kDa molecule by immunoblotting, and its location was detected on the M. tuberculosis surface by transmission electron microscopy, suggesting that it is a mycobacterial surface protein. Binding assays led to recognizing high activity binding peptides (HABP); five HABPs specifically bound to U937 cells, and six specifically bound to A549 cells. HABP circular dichroism suggested that they had an α-helical structure. HABP-target cell interaction was determined to be specific and saturable; some of them also displayed greater affinity for A549 cells than U937 cells. The critical amino acids directly involved in their interaction with U937 cells were also determined. Two probable receptor molecules were found on U937 cells and five on A549 for the two HABPs analyzed. These observations have important biological significance for studying bacillus-target cell interactions and implications for developing strategies for controlling this disease. Copyright {\circledC} 2005 The Protein Society.",
author = "Martha Forero and {\'A}lvaro Puentes and Jimena Cort{\'e}s and Fabio Castillo and Ricardo Vera and Rodr{\'i}guez, {Luis E.} and John Valbuena and Marisol Ocampo and Hernando Curtidor and Jaiver Rosas and Javier Garc{\'i}a and Gloria Barrera and Rosalba Alfonso and Patarroyo, {Manuel A.} and Patarroyo, {Manuel E.}",
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Forero, M, Puentes, Á, Cortés, J, Castillo, F, Vera, R, Rodríguez, LE, Valbuena, J, Ocampo, M, Curtidor, H, Rosas, J, García, J, Barrera, G, Alfonso, R, Patarroyo, MA & Patarroyo, ME 2005, 'Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells', Protein Science, pp. 2767-2780. https://doi.org/10.1110/ps.051592505

Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells. / Forero, Martha; Puentes, Álvaro; Cortés, Jimena; Castillo, Fabio; Vera, Ricardo; Rodríguez, Luis E.; Valbuena, John; Ocampo, Marisol; Curtidor, Hernando; Rosas, Jaiver; García, Javier; Barrera, Gloria; Alfonso, Rosalba; Patarroyo, Manuel A.; Patarroyo, Manuel E.

In: Protein Science, 01.11.2005, p. 2767-2780.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identifying putative Mycobacterium tuberculosis Rv2004c protein sequences that bind specifically to U937 macrophages and A549 epithelial cells

AU - Forero, Martha

AU - Puentes, Álvaro

AU - Cortés, Jimena

AU - Castillo, Fabio

AU - Vera, Ricardo

AU - Rodríguez, Luis E.

AU - Valbuena, John

AU - Ocampo, Marisol

AU - Curtidor, Hernando

AU - Rosas, Jaiver

AU - García, Javier

AU - Barrera, Gloria

AU - Alfonso, Rosalba

AU - Patarroyo, Manuel A.

AU - Patarroyo, Manuel E.

PY - 2005/11/1

Y1 - 2005/11/1

N2 - Virulence and immunity are still poorly understood in Mycobacterium tuberculosis. The H37Rv M. tuberculosis laboratory strain genome has been completely sequenced, and this along with proteomic technology represent powerful tools contributing toward studying the biology of target cell interaction with a facultative bacillus and designing new strategies for controlling tuberculosis. Rv2004c is a putative M. tuberculosis protein that could have specific mycobacterial functions. This study has revealed that the encoding gene is present in all mycobacterium species belonging to the M. tuberculosis complex. Rv2004c gene transcription was observed in all of this complex's strains except Mycobacterium bovis and Mycobacterium microti. Rv2004c protein expression was confirmed by using antibodies able to recognize a 54-kDa molecule by immunoblotting, and its location was detected on the M. tuberculosis surface by transmission electron microscopy, suggesting that it is a mycobacterial surface protein. Binding assays led to recognizing high activity binding peptides (HABP); five HABPs specifically bound to U937 cells, and six specifically bound to A549 cells. HABP circular dichroism suggested that they had an α-helical structure. HABP-target cell interaction was determined to be specific and saturable; some of them also displayed greater affinity for A549 cells than U937 cells. The critical amino acids directly involved in their interaction with U937 cells were also determined. Two probable receptor molecules were found on U937 cells and five on A549 for the two HABPs analyzed. These observations have important biological significance for studying bacillus-target cell interactions and implications for developing strategies for controlling this disease. Copyright © 2005 The Protein Society.

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DO - 10.1110/ps.051592505

M3 - Article

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JF - Protein Science

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