Protein profiles of bacteriophages of the family Myoviridae-like induced on M. haemolytica

Abstract

The aim of study was to isolate, characterize and analyse the protein profiles of Myoviridae-like bacteriophages obtained from M. haemolytica using MALDI TOF mass spectrometry. The material consisted of the M. haemolytica reference strain ATCC® BAA410, reference serotypes A1, A2, A5, A6, A7, A9, and A11, and wild-type isolates of serotype A1. Bacteriophage morphology was examined with a transmission electron microscope. The proteins were separated in SDS-PAGE and two-dimensional electrophoresis and characterized by MALDI-TOF. Among the phages obtained, seven were specific for strains A1, A2, A5, A6, A7 and 25, and PHL-1 was specific for the BAA410 strain. The protein profiles for the phages were very similar to one another, but differed from the reference phage in that they lacked protein fractions with molecular weights of 22.9, 56.3 and 73.1 kDa. 2D electrophoresis revealed significant differences in the size of proteins and their localization in the pH gradient. The most similar profiles were observed in phages specific for strains BAA-410 and A6. In all profiles two main spots were observed in the molecular weight range from 44 to 70 kDa at pH < 4. The results indicate that 2D electrophoresis is a very useful tool for characterization of phage protein profiles. An important objective was to determine the molecular differences between morphologically similar phages belonging to one family and to find similarities to phages specific for other pathogens. The study also assessed the suitability of the methods used to characterize phages.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0630-3) contains supplementary material, which is available to authorized users.

Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo

“…Several reports have investigated the structural protein proteome, but there are few studies on total protein profiles of phage. Previously 2D electrophoresis coupled with MALDI-TOF was used to investigate the total proteins of a Myoviridae phage specific for Mannheimia haemolytica 41 , while a total of 346 proteins of phage BPA43 specific for ESBL-KP were characterized by shotgun proteomics 42 . In this study shotgun proteomic analysis combined with bioinformatics analysis of phage KP1801 identified 323 proteins.…”

Virulent Drexlervirial Bacteriophage MSK, Morphological and Genome Resemblance With Rtp Bacteriophage Inhibits the Multidrug-Resistant Bacteria

“…Further, the structure profile of bacteriophages is determined by SDS-PAGE, enabling us to determine the individual protein, molecular weight, and size. These analyses can be used for the morphotyping and differentiation of distinct proteins even within the same species ( Urban-Chmiel et al, 2018 ). ORF prediction revealed that fewer genes are encoding structural proteins.…”

The Analysis of OmpA and Rz/Rz1 of Lytic Bacteriophage from Surabaya, Indonesia

“…The isolation phase of bacteriophage protein began with a suspension of 0.5 mL supplemented with 0.5 mL of 96% EtOH and then incubated at −20°C for 24 hours. 100 μ L of single-power SDS-PAGE sample lysis buffer was added to the protein precipitation following the separation of ethanol layer (Tris-HCl 62,5 m, 2% SDS, 6% 2-mercaptoethanol, 10% glycerol, 0,1% blue bromophenol, pH 6,8) and then heated to 10 minutes at 100°C [ 23 ].…”