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Characterizing the Mycobacterium tuberculosis Rv2707 protein and determining its sequences which specifically bind to two human cell lines

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Chapeton?Montes, Julie A.
Plaza, David F.
Curtidor, Hernando
Forero, Martha
Vanegas, Magnolia
Patarroyo, Manuel E.
Patarroyo, Manuel A.

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2008

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Abstract
The Rv2707 gene encoding a putative alanine- and leucine-rich protein was found to be present in all Mycobacterium tuberculosis complex strains (by PCR) and its transcription was shown by RT-PCR in all but M. bovis and M. microti. Antibodies raised against Rv2707 peptides specifically recognized the native protein by Western blot and were able to locate this protein on the M. tuberculosis membrane by immunoelectron microscopy. A549 and U937 cells lines were used in binding assays involving synthetic peptides covering the whole Rv2707 protein. High A549 cell-binding peptide 16083 ( 281QEEWPAPATHAHRLGNWLKAY300) was identified. Peptides 16072 (61LFGPDTLPAIEKSALSTAHSY80) and 16084 ( 301RIGVGTTTYSSTAQHSAVAA320) presented high specific binding to both A549 and U937 cells. Cross-linking assays revealed that peptide 16084 specifically bound to a 40-kDa and a 50-kDa U937 cell membrane protein. High activity binding peptides (HABPs) 16083 and 16084 were able to inhibit M. tuberculosis invasion of A549 cells. Our results suggest that these sequences could be part of the binding sites used by the bacillus for interacting with target cells, and thus represent good candidates to be tested in a future subunit-based, multiepitope, antituberculosis vaccine. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 The Protein Society.
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Bacterial protein , Protein rv2707 , Unclassified drug , Amino acid sequence , Article , Controlled study , Gene location , Gene sequence , Human , Human cell , Mycobacterium tuberculosis , Nonhuman , Priority journal , Reverse transcription polymerase chain reaction , Amino acid sequence , Bacterial proteins , Cell line , Computational biology , Humans , Microscopy , Molecular sequence data , Mycobacterium tuberculosis , Peptide fragments , Protein binding , U937 cells , Mycobacterium tuberculosis , Mycobacterium tuberculosis complex , A549 cell , High activity binding peptide (habp) , Invasion , Mycobacterium tuberculosis , Rv2707 , U937 cell
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