The Mycobacterium tuberculosis membrane protein Rv2560 − biochemical and functional studies

Abstract

Tuberculosis (TB) is a major public health problem throughout the world, affecting almost nine million people [1] and causing more than three million deaths per year. An increasing incidence of TB, related to the high risk of developing the disease in immunosuppressed individuals and the increasing proportion of Mycobacterium tuberculosis drug-resistant strains, has contributed to this problem [2,3]. This makes the Keywords high-activity binding peptide; invasion inhibition; Mycobacterium tuberculosishost cell interaction; Rv2560 membrane protein Correspondence M. E. Patarroyo, Carrera 50 # 26-00,

Mycobacterium tuberculosis Rv0679c protein sequences involved in host-cell infection: Potential TB vaccine candidate antigen

“…A highly specific binding assay was used to evaluate ligand-receptor interactions established between Rv0679c peptides and A549 and U937 cell surface receptors, same as has been reported for other mycobacterial proteins [23-25,37]. Based on this methodology, two HABPs binding with high activity to both cell lines were identified (namely HABPs 30979 and 30987), while other two HABPs (30985 and 30986) bound only to A549 cells.…”

“…Any peptide displaying a specific binding activity of ≥1% was considered a HABP [23-25,37]. Binding constants were determined by performing a saturation assay using U937 cells and peptide concentrations larger than the ones used for binding assays (0-4500 nM).…”

Peptides derived from Mycobacterium tuberculosis Rv2301 protein are involved in invasion to human epithelial cells and macrophages

“…interaction and complement previous studies reported by our group (Chapeton-Montes et al 2008b;Plaza et al 2007;Vera-Bravo et al 2005). The same highly sensitive and robust methodology has been used to identify and characterize amino acid sequences mediating binding interactions with target cells, and that have the ability to inhibit mycobacterial invasion.…”

“…The possible biological importance of the HABPs identified in Rv2301 in mediating Mycobacterium-host cell interactions was analyzed according to a methodology previously reported by our institute (Chapeton-Montes et al 2008a, b;Plaza et al 2007;Vera-Bravo et al 2005). As shown in Fig.…”

“…After incubation, unbound peptides were removed from cells by sedimentation through a dibutylphthalate-dioctylphthalate cushion mixture (d = 1.015 g/mL) and centrifuged at 9,000g for 2 min Plaza et al 2007;Vera-Bravo et al 2005). Cell-bound peptides were measured in an automatic gamma counter.…”