Abstract
Abstract The aim of this study was to examine the protein expression profiles of persistentChlamydia pneumoniae by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Although 2D PAGE is still the method of choice for separating and detecting components of complex protein mixtures, it has several distinct disadvantages; i.e., being labor-intensive and having a bias toward proteins within the dynamic range of the gel condition. Hence, SELDI-TOF-MS technology was used to complement 2D PAGE.C. pneumoniae-infected HEp2 cells were treated with or without IFN-γ, and protein expression profiles were determined at 48 h postinfection (hpi). Unfractionated monolayers were also used for protein profiling by SELDI-TOF, using two different chip surface types: weak cation exchanger and hydrophobic surface. Under IFN-γ-induced persistence,C. pneumoniae expresses an altered protein expression profile. Twenty chlamydial proteins showed differential regulatory patterns by SELDI-TOF-MS, two of which, HSP-70 cofactor, and a hypothetical protein, were identified by 2D PAGE and mass spectrometry. Two additional proteins, phosphatidylserine decarboxylase and 30S ribosomal protein S17, were exclusively identified by SELDI TOF-MS analysis, as these were not present in sufficient quantity for detection by 2D PAGE. We propose that a combination of 2D-PAGE and SELDI-TOF-MS may complement the disadvantages of each technique alone and may provide a rapid and precise screening technique.
Highlights
Chlamydia pneumoniae is a Gram-negative obligate intracellular pathogen which has a complex biphasic developmental cycle and causes both acute and chronic respiratory infections
We propose that a combination of 2D-PAGE and SELDI-TOF-MS may complement the disadvantages of each technique alone and may provide a rapid and precise screening technique
The reticulate bodies (RB) divides by binary fission and subsequently redifferentiates to elementary bodies (EB), and are released from the host cell to initiate another lytic cycle [6,7,8,9,10]
Summary
Chlamydia pneumoniae is a Gram-negative obligate intracellular pathogen which has a complex biphasic developmental cycle and causes both acute and chronic respiratory infections. It has been implicated in chronic human disease, atherosclerosis [1,2,3,4,5]. A subinhibitory concentration of IFN-γ can lead to a state in which redifferentiation of RB to infectious EB is interrupted and the RB develop into an arrested growth stage, commonly known as an aberrant body (AB) Even though it maintains viability while inside the host cell, it is non-culturable and results in a long-term relationship within the host cells [11,12]
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