The Mycobacterium tuberculosis membrane protein Rv2560 - Biochemical and functional studies

David F. Plaza, Hernando Curtidor, Manuel A. Patarroyo, Julie A. Chapeton-Montes, Claudia Reyes, Jose Barreto, Manuel E. Patarroyo

Resultado de la investigación: Contribución a RevistaArtículo

12 Citas (Scopus)

Resumen

The characterization of membrane proteins having no identified function in Mycobacterium tuberculosis is important for a better understanding of the biology of this pathogen. In this work, the biological activity of the Rv2560 protein was characterized and evaluated. Primers used in PCR and RT-PCR assays revealed that the gene encoding protein Rv2560 is present in M. tuberculosis complex strains, but transcribed in only some of them. Sera obtained from rabbits inoculated with polymer peptides from this protein recognized a 33 kDa band in the M. tuberculosis lysate and a membrane fraction corresponding to the predicted molecular mass (33.1 kDa) of this protein. Immunoelectron microscopy analysis found this protein on the mycobacterial membrane. Sixteen peptides covering its entire length were chemically synthesized and tested for their ability to bind to A549 and U937 cells. Peptide 11024 (121VVALSDRATTAYTNTSGVSS140) showed high specific binding to both cell types (dissociation constants of 380 and 800 nm, respectively, and positive receptor-ligand interaction cooperativity), whereas peptide 11033 (284LIGIPVAALIHVYTYRKLSGG304) displayed high binding activity to A549 cells only. Cross-linking assays showed the specific binding of peptide 11024 to a 54 kDa membrane protein on U937. Invasion inhibition assays, in the presence of shared high-activity binding peptide identified for U937 and A549 cells, presented maximum inhibition percentages of 50.53% and 58.27%, respectively. Our work highlights the relevance of the Rv2560 protein in the M. tuberculosis invasion process of monocytes and epithelial cells, and represents a fundamental step in the rational selection of new antigens to be included as components in a multiepitope, subunit-based, chemically synthesized, antituberculosis vaccine. © 2007 The Authors.
Idioma originalEnglish (US)
Páginas (desde-hasta)6352-6364
Número de páginas13
PublicaciónFEBS Journal
DOI
EstadoPublished - dic 1 2007

Huella dactilar

Mycobacterium tuberculosis
Membrane Proteins
Peptides
Assays
U937 Cells
Proteins
Membranes
Polymerase Chain Reaction
Gene encoding
Immunoelectron Microscopy
Molecular mass
Pathogens
Bioactivity
Monocytes
Microscopic examination
Polymers
Vaccines
Epithelial Cells
Rabbits
Ligands

Citar esto

Plaza, D. F., Curtidor, H., Patarroyo, M. A., Chapeton-Montes, J. A., Reyes, C., Barreto, J., & Patarroyo, M. E. (2007). The Mycobacterium tuberculosis membrane protein Rv2560 - Biochemical and functional studies. FEBS Journal, 6352-6364. https://doi.org/10.1111/j.1742-4658.2007.06153.x
Plaza, David F. ; Curtidor, Hernando ; Patarroyo, Manuel A. ; Chapeton-Montes, Julie A. ; Reyes, Claudia ; Barreto, Jose ; Patarroyo, Manuel E. / The Mycobacterium tuberculosis membrane protein Rv2560 - Biochemical and functional studies. En: FEBS Journal. 2007 ; pp. 6352-6364.
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abstract = "The characterization of membrane proteins having no identified function in Mycobacterium tuberculosis is important for a better understanding of the biology of this pathogen. In this work, the biological activity of the Rv2560 protein was characterized and evaluated. Primers used in PCR and RT-PCR assays revealed that the gene encoding protein Rv2560 is present in M. tuberculosis complex strains, but transcribed in only some of them. Sera obtained from rabbits inoculated with polymer peptides from this protein recognized a 33 kDa band in the M. tuberculosis lysate and a membrane fraction corresponding to the predicted molecular mass (33.1 kDa) of this protein. Immunoelectron microscopy analysis found this protein on the mycobacterial membrane. Sixteen peptides covering its entire length were chemically synthesized and tested for their ability to bind to A549 and U937 cells. Peptide 11024 (121VVALSDRATTAYTNTSGVSS140) showed high specific binding to both cell types (dissociation constants of 380 and 800 nm, respectively, and positive receptor-ligand interaction cooperativity), whereas peptide 11033 (284LIGIPVAALIHVYTYRKLSGG304) displayed high binding activity to A549 cells only. Cross-linking assays showed the specific binding of peptide 11024 to a 54 kDa membrane protein on U937. Invasion inhibition assays, in the presence of shared high-activity binding peptide identified for U937 and A549 cells, presented maximum inhibition percentages of 50.53{\%} and 58.27{\%}, respectively. Our work highlights the relevance of the Rv2560 protein in the M. tuberculosis invasion process of monocytes and epithelial cells, and represents a fundamental step in the rational selection of new antigens to be included as components in a multiepitope, subunit-based, chemically synthesized, antituberculosis vaccine. {\circledC} 2007 The Authors.",
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The Mycobacterium tuberculosis membrane protein Rv2560 - Biochemical and functional studies. / Plaza, David F.; Curtidor, Hernando; Patarroyo, Manuel A.; Chapeton-Montes, Julie A.; Reyes, Claudia; Barreto, Jose; Patarroyo, Manuel E.

En: FEBS Journal, 01.12.2007, p. 6352-6364.

Resultado de la investigación: Contribución a RevistaArtículo

TY - JOUR

T1 - The Mycobacterium tuberculosis membrane protein Rv2560 - Biochemical and functional studies

AU - Plaza, David F.

AU - Curtidor, Hernando

AU - Patarroyo, Manuel A.

AU - Chapeton-Montes, Julie A.

AU - Reyes, Claudia

AU - Barreto, Jose

AU - Patarroyo, Manuel E.

PY - 2007/12/1

Y1 - 2007/12/1

N2 - The characterization of membrane proteins having no identified function in Mycobacterium tuberculosis is important for a better understanding of the biology of this pathogen. In this work, the biological activity of the Rv2560 protein was characterized and evaluated. Primers used in PCR and RT-PCR assays revealed that the gene encoding protein Rv2560 is present in M. tuberculosis complex strains, but transcribed in only some of them. Sera obtained from rabbits inoculated with polymer peptides from this protein recognized a 33 kDa band in the M. tuberculosis lysate and a membrane fraction corresponding to the predicted molecular mass (33.1 kDa) of this protein. Immunoelectron microscopy analysis found this protein on the mycobacterial membrane. Sixteen peptides covering its entire length were chemically synthesized and tested for their ability to bind to A549 and U937 cells. Peptide 11024 (121VVALSDRATTAYTNTSGVSS140) showed high specific binding to both cell types (dissociation constants of 380 and 800 nm, respectively, and positive receptor-ligand interaction cooperativity), whereas peptide 11033 (284LIGIPVAALIHVYTYRKLSGG304) displayed high binding activity to A549 cells only. Cross-linking assays showed the specific binding of peptide 11024 to a 54 kDa membrane protein on U937. Invasion inhibition assays, in the presence of shared high-activity binding peptide identified for U937 and A549 cells, presented maximum inhibition percentages of 50.53% and 58.27%, respectively. Our work highlights the relevance of the Rv2560 protein in the M. tuberculosis invasion process of monocytes and epithelial cells, and represents a fundamental step in the rational selection of new antigens to be included as components in a multiepitope, subunit-based, chemically synthesized, antituberculosis vaccine. © 2007 The Authors.

AB - The characterization of membrane proteins having no identified function in Mycobacterium tuberculosis is important for a better understanding of the biology of this pathogen. In this work, the biological activity of the Rv2560 protein was characterized and evaluated. Primers used in PCR and RT-PCR assays revealed that the gene encoding protein Rv2560 is present in M. tuberculosis complex strains, but transcribed in only some of them. Sera obtained from rabbits inoculated with polymer peptides from this protein recognized a 33 kDa band in the M. tuberculosis lysate and a membrane fraction corresponding to the predicted molecular mass (33.1 kDa) of this protein. Immunoelectron microscopy analysis found this protein on the mycobacterial membrane. Sixteen peptides covering its entire length were chemically synthesized and tested for their ability to bind to A549 and U937 cells. Peptide 11024 (121VVALSDRATTAYTNTSGVSS140) showed high specific binding to both cell types (dissociation constants of 380 and 800 nm, respectively, and positive receptor-ligand interaction cooperativity), whereas peptide 11033 (284LIGIPVAALIHVYTYRKLSGG304) displayed high binding activity to A549 cells only. Cross-linking assays showed the specific binding of peptide 11024 to a 54 kDa membrane protein on U937. Invasion inhibition assays, in the presence of shared high-activity binding peptide identified for U937 and A549 cells, presented maximum inhibition percentages of 50.53% and 58.27%, respectively. Our work highlights the relevance of the Rv2560 protein in the M. tuberculosis invasion process of monocytes and epithelial cells, and represents a fundamental step in the rational selection of new antigens to be included as components in a multiepitope, subunit-based, chemically synthesized, antituberculosis vaccine. © 2007 The Authors.

U2 - 10.1111/j.1742-4658.2007.06153.x

DO - 10.1111/j.1742-4658.2007.06153.x

M3 - Article

SP - 6352

EP - 6364

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

ER -