Comparative Proteome Analysis of the Outer Membrane Proteins of in Vitro‐Induced Multi‐Drug Resistant Neisseria gonorrhoeae

Abstract

Antimicrobial-resistant gonococcus has been a major problem in sexually transmitted disease control. Outer membrane proteins (OMPs) of Neisseria gonorrhoeae were suggested to have influence on its resistance to antibiotics. So, in this work, we provide a proteomic analysis tool for examining the OMPs of N. gonorrhoeae and also provide a comparative analysis of the OMPs between the susceptible parent strain (92WT) and the resistance-induced isogenic mutant (92mu13) to determine the OMPs responsible for resistance. The 2-D gel spots of 92mu13 differed from 92WT particularly in porin, pilus secretion protein (PilQ) and enzymes. PilQ expression in 92mu13 was considerably reduced by abrupt termination at nucleotide 2,112. This made it difficult to form a high molecular mass (HMM) pore at the outer membrane; it is suspected that reduction of PilQ serves a role in antibiotic resistance in N. gonorrhoeae. The amount of porin was not changed but its isoelectric point (pI) shifted to a basic region, which is caused by the alteration of an amino acid of porin and it is suggested to relate to the development of antimicrobial resistance. Differential regulation of the enzymes involved in metabolism was found in 92mu13, believed to represent an adaptation of N. gonorrhoeae to the antibiotic environment.

Quantitative Proteomics of the 2016 WHO Neisseria gonorrhoeae Reference Strains Surveys Vaccine Candidates and Antimicrobial Resistance Determinants

“…In addition, subcellular fractionation steps preceding proteomic applications reduce sample complexity, increase the likelihood of discovering low-abundance proteins, and aid in defining protein localization, all of which provide further insights into the proteins' functions and interactomes (43,44). For N. gonorrhoeae, proteomic approaches have begun to deliver proteinaceous vaccine candidates (29,30,39,45) and to support elucidation of AMR patterns (46,47). Current off-gel proteomics, such as isobaric tag labeling (isobaric tagging for absolute quantification, iTRAQ; and tandem mass tags, TMT) coupled with highpressure liquid chromatography and mass spectrometry techniques (LC-MS/MS), demonstrate superb protein separation and identification and enable detection of proteins in the low femtomole to high attomole range with precision and reliability (29,48,49).…”

Proteomic identification of membrane proteins regulating antimicrobial peptide resistance inVibrio parahaemolyticus

“…2008). Comparative proteome analyses have also been performed to investigate multi‐drug–resistant Neisseria gonorrhoeae (Yoo et al. 2007) and vancomycin‐resistant Staphylococcus aureus (Pieper et al.…”

“…Peng and colleagues have successfully identified a network of outer membrane proteins (OMPs) regulating antibiotic resistance in Escherichia coli and Pseudomonas aeruginosa, using proteomic approaches (Peng et al 2005;Xu et al 2006;Li et al 2007Lin et al 2008). Comparative proteome analyses have also been performed to investigate multi-drug-resistant Neisseria gonorrhoeae (Yoo et al 2007) and vancomycin-resistant Staphylococcus aureus (Pieper et al 2006;Drummelsmith et al 2007). In the present study, we seek to identify proteins regulating AMP resistance in V. parahaemolyticus, using membrane subproteome analysis.…”

Quantitative Proteomics of the Neisseria Gonorrhoeae Cell Envelope and Membrane Vesicles for the Discovery of Potential Therapeutic Targets

“…Although the potential importance of proteins localized to the GC cell envelope and MVs has been reported previously (25,26), only two proteomic studies have been published addressing GC membrane composition (27,28). Most studies have focused on extensive characterization of factors involved in direct host cell interaction: protruding surface proteins (pili), outer membrane adhesins Opa, porins P.IA and P.IB, lipooligosaccharide, and several iron utilization proteins (3, 4, 15, 29 -32).…”