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Protein expression analysis ofChlamydia pneumoniae persistence by combined surface-enhanced laser desorption ionization time-of-flight mass spectrometry and two-dimensional polyacrylamide gel electrophoresis

Clinical Proteomics volume 2, pages 205–215 (2006)Cite this article

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.

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Authors and Affiliations

  1. Division of Infectious Diseases, Department of Medicine, University of Louisville School of Medicine, Louisville, KY

    Sanghamitra Mukhopadhyay, Erin D. Sullivan & James T. Summersgill

  2. Department of Pathology and Laboratory Medicine, University of Louisville School of Medicine, Louisville, KY

    Saeed A. Jortani, Roland Valdes & Christine Q. Simmons

  3. Department of Microbiology and Immunology, University of Louisville School of Medicine, Louisville, KY

    Richard D. Miller & James T. Summersgill

  4. Biological Division Directorate, Naval Medical Research Center, Rockville, MD

    James T. Summersgill

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  1. Sanghamitra Mukhopadhyay
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  2. Saeed A. Jortani
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  3. Roland Valdes
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  4. Christine Q. Simmons
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Correspondence to James T. Summersgill.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Mukhopadhyay, S., Jortani, S.A., Valdes, R. et al. Protein expression analysis ofChlamydia pneumoniae persistence by combined surface-enhanced laser desorption ionization time-of-flight mass spectrometry and two-dimensional polyacrylamide gel electrophoresis. Clin Proteom 2, 205–215 (2006). https://doiorg.publicaciones.saludcastillayleon.es/10.1007/BF02752501

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Clinical Proteomics

ISSN: 1559-0275