Proteomic identification of responsive proteins of Vibrio parahaemolyticus under high hydrostatic pressure.
BACKGROUND
High hydrostatic pressure (HHP) processing is currently being used as a treatment for certain foods to inhibit spoilage organisms and control the presence of foodborne pathogens. In this study proteome profiles were performed by two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF/TOF identification to determine the effects of HHP (50, 100, 150 and 200 MPa, each for 10 min) on Vibrio parahaemolyticus ATCC 17802 (∼8 log CFU mL⁻¹) in order to understand how it responds to mechanical stress injury.


RESULTS
Multiple comparisons of 2-DE revealed that the majority of changes in protein abundance occurred in a pressure-dependent fashion. A total of 18 differentially expressed protein spots were successfully identified by MALDI-TOF/TOF analysis. Moreover, quantitative RT-PCR and immunoblotting also substantiated the changes of transcriptional and translational levels of representative proteins.


CONCLUSIONS
Our results suggested that V. parahaemolyticus may respond to HHP treatment through suppression of membrane stability and functionality (PfaC, Alr2, MltA, PLA2 and PatH), depression of biosynthesis and cellular processes (NadB, PyrB and ArgB), decreased levels of transcription (RpoD) and translation (RpsA, RplJ and PheS), and effective activation of protein folding and stress-related elements (GroES, DnaK and GroEL). This study may provide insight into the nature of the cellular targets of high pressure and in high-pressure resistance mechanisms in V. parahaemolyticus.
Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310035, P.R. China.
Ling-Lin
LL
Fu
Rui
R
Wang
Yanbo
Y
Wang
Junda
J
Lin
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D030562:Databases, Protein; D005519:Food Preservation / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial; D006874:Hydrostatic Pressure; D050296:Microbial Viability; D046529:Microscopy, Electron, Transmission; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013312:Stress, Physiological; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis; D014736:Vibrio parahaemolyticus / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure
D001426:Bacterial Proteins; D020543:Proteome
Vibrio parahaemolyticus; high hydrostatic pressure (HHP); proteomics; responsive protein