Expression, purification and crystallization of phosphoribosyl transferase from a mycobacteriophage

Abstract

Tuberculosis (TB) continues to remain a leading cause of death globally. Of particular concern is the emergence and rise in incidence of multidrug-resistant and extremely drug-resistant cases of TB. To counter this threat, it is important to explore alternative therapies, including phage therapy. Phage BTCU-1 specifically infects Mycobacterium spp. and kills the majority of them. Intriguingly, many proteins from the phage do not share high amino-acid sequence identity with proteins from species other than phages. Here, the expression, purification and crystallization of one such protein, a putative phosphoribosyl transferase from phage BTCU-1, is reported. The crystals belonged to space group C222, with unit-cell parameters a = 59.71, b = 64.42, c = 65.32 Å, α = β = γ = 90°. The crystals diffracted X-rays to 2.2 Å resolution.

<p>Phage therapy as a renewed therapeutic approach to mycobacterial infections: a comprehensive review</p>

“…Mycobacteriophage BTCU-1 belongs to Siphoviridae family and has been isolated from soil specimen, obtained from eastern Taiwan 80. The length of the mycobacteriophage BTCU-1 genome is approximately 46 kb, and this bacteriophage has a linear double-stranded DNA with an icosahedral head and a very long tail 81.…”

“…The genome of BTCU-1 encodes several proteins with predefined functions. One of these proteins is identified as a putative phosphoribosyl transferase (PRT) and is particularly found in mycobacteriophages that infect Mycobacterium 80,82. These proteins eradicate most bacteria during the course of their life cycle 80.…”

“…One of these proteins is identified as a putative phosphoribosyl transferase (PRT) and is particularly found in mycobacteriophages that infect Mycobacterium 80,82. These proteins eradicate most bacteria during the course of their life cycle 80. Moreover, different mycobacteriophages encode lytic endolysins that have antimicrobial activity and can be effective against MDR and XDR MTB strains 83–85.…”