Single Amino Acid Change Mutation in the Hydrophobic Core of the N-terminal Domain of P22 TSP affects the Proteins Stability

Abstract

The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has significantly shifted the attention of researchers to critically investigate most viruses to understand specific characteristics that impart their virulence. For instance, the SARS-CoV-2 has undergone several mutations, with some variants classified as variants of concern, e.g., the Omicron and Delta variant of SARS-CoV-2 are known for their rapid transmission and antigenicity due to mutation in the Spike protein. P22 bacteriophage is a bacterial virus that has a tailspike protein (TSP) that performs similar functions as the Spike protein of SARS-COV-2. We previously carried out a site-directed mutagenesis of the P22 TSP to bear disruptive mutations in the hydrophobic core of the N-terminal Domain (NTD), then partially characterized the properties of the mutant TSPs. In this process, the valine patch (triple valine residues that formed a hydrophobic core) was replaced with charged amino acids (Asp or lysine) or hydrophobic amino acids (Leucine or isoleucine). Some of the mutant TSPs characterized showed significant differences in migration in both native and SDS-PAGE. Mutants with such disruptive mutation are known to show non-native properties, and as expected, most of these mutants obtained showed significantly different properties from the WT P22 TSP. In this work, we further characterized these mutant species by computational and in vitro assays to demonstrate the validity of our previous inference that the valine patch is a critical player in the stability of the N-terminal domain of the P22 TSP.

Recent trends in the use of bacteriophages as replacement of antimicrobials against food-animal pathogens

“…On glass Petri plates, C. jejuni biofilm formation was prevented using the lytic bacteriophages CP8 and CP30 that were isolated from chicken excrement ( 89 ). In comparison to a control biofilm that had not been treated with bacteriophage, The viable count was found to be reduced by 1.0–3.0 log CFU/cm2 24 h after infection following phage treatment of each biofilm, according to the research depending on the strain that forms a biofilm, there were differences in the degree of reduction and the potential for phage resistance ( 90 ). When compared to C. jejuni strain NCTC 11168, C. jejuni strain PT14 was distinguished by its ability to produce a much larger amount of biofilm on glass.…”

Proteolytic Analysis of Epsilon 34 Phage Tailspike protein indicates Partial Sensitivity to Proteinase K

“…The of ε34 phage has been demonstrated to possesses similar functional and structural characteristics as the P22 phage TSP [10] . The predicted structure of Epsilon 34 TSP consists of a globular head binding domain, a solenoid-shape parallel beta-helix domain that is used in the LPS binding process as also found in the P22 phage TSP [9, 14, 15] . Phages or phage derived components can complement host cell defense against bacteria [3, 16] .…”