Characterization of a secreted Chlamydia protease

Allan C Shaw, Brian Vandahl, Martin Røssel Larsen, Peter Roepstorff, Kris Gevaert, Joël Vandekerckhove, Gunna Christiansen, Svend Birkelund

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Chlamydiae are obligate intracellular bacteria that are important human pathogens. The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified. Most likely, effector proteins are secreted in order to promote infection. Effector proteins cannot be identified by motif or similarity searches. As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of [35S]-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified Chlamydia. Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method. Two protein spots were identified among the candidates. These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D-PAGE profiles of whole lysates of infected cells but absent from purified Chlamydia. CPAF was recently identified by Zhong and colleagues as a secreted protease which cleaves host cell transcription factors essential for MHC class I and II antigen presentation. The identification of CPAF in this paper verifies the applicability of the described method for the identification of secreted proteins. We extend the findings by Zhong et al. by proteome studies of expression and turnover of C. trachomatis CPAF showing that the degradation of C. trachomatis D CPAF in the host cell is very limited. Furthermore, we show that two fragments of CPAF exist in C. pneumoniae as well as in C. trachomatis.
OriginalsprogEngelsk
TidsskriftCellular Microbiology
Vol/bind4
Udgave nummer7
Sider (fra-til)411-24
ISSN1462-5814
StatusUdgivet - 2002

Fingeraftryk

Peptide Hydrolases
Chlamydia trachomatis
Proteins
Chlamydophila pneumoniae
Antigen Presentation
Proteome

Citer dette

Shaw, A. C., Vandahl, B., Larsen, M. R., Roepstorff, P., Gevaert, K., Vandekerckhove, J., ... Birkelund, S. (2002). Characterization of a secreted Chlamydia protease. Cellular Microbiology, 4(7), 411-24.
Shaw, Allan C ; Vandahl, Brian ; Larsen, Martin Røssel ; Roepstorff, Peter ; Gevaert, Kris ; Vandekerckhove, Joël ; Christiansen, Gunna ; Birkelund, Svend. / Characterization of a secreted Chlamydia protease. I: Cellular Microbiology. 2002 ; Bind 4, Nr. 7. s. 411-24.
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abstract = "Chlamydiae are obligate intracellular bacteria that are important human pathogens. The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified. Most likely, effector proteins are secreted in order to promote infection. Effector proteins cannot be identified by motif or similarity searches. As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of [35S]-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified Chlamydia. Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method. Two protein spots were identified among the candidates. These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D-PAGE profiles of whole lysates of infected cells but absent from purified Chlamydia. CPAF was recently identified by Zhong and colleagues as a secreted protease which cleaves host cell transcription factors essential for MHC class I and II antigen presentation. The identification of CPAF in this paper verifies the applicability of the described method for the identification of secreted proteins. We extend the findings by Zhong et al. by proteome studies of expression and turnover of C. trachomatis CPAF showing that the degradation of C. trachomatis D CPAF in the host cell is very limited. Furthermore, we show that two fragments of CPAF exist in C. pneumoniae as well as in C. trachomatis.",
keywords = "Amino Acid Sequence, Antibodies, Bacterial, Base Sequence, Cell Line, Chlamydia trachomatis, Chlamydophila pneumoniae, Cloning, Molecular, DNA, Bacterial, Electrophoresis, Gel, Two-Dimensional, Endopeptidases, Hela Cells, Humans, Microscopy, Fluorescence, Molecular Sequence Data, Peptide Fragments, Protease Inhibitors, Species Specificity, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization",
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Shaw, AC, Vandahl, B, Larsen, MR, Roepstorff, P, Gevaert, K, Vandekerckhove, J, Christiansen, G & Birkelund, S 2002, 'Characterization of a secreted Chlamydia protease', Cellular Microbiology, bind 4, nr. 7, s. 411-24.

Characterization of a secreted Chlamydia protease. / Shaw, Allan C; Vandahl, Brian; Larsen, Martin Røssel; Roepstorff, Peter; Gevaert, Kris; Vandekerckhove, Joël; Christiansen, Gunna; Birkelund, Svend.

I: Cellular Microbiology, Bind 4, Nr. 7, 2002, s. 411-24.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Characterization of a secreted Chlamydia protease

AU - Shaw, Allan C

AU - Vandahl, Brian

AU - Larsen, Martin Røssel

AU - Roepstorff, Peter

AU - Gevaert, Kris

AU - Vandekerckhove, Joël

AU - Christiansen, Gunna

AU - Birkelund, Svend

PY - 2002

Y1 - 2002

N2 - Chlamydiae are obligate intracellular bacteria that are important human pathogens. The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified. Most likely, effector proteins are secreted in order to promote infection. Effector proteins cannot be identified by motif or similarity searches. As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of [35S]-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified Chlamydia. Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method. Two protein spots were identified among the candidates. These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D-PAGE profiles of whole lysates of infected cells but absent from purified Chlamydia. CPAF was recently identified by Zhong and colleagues as a secreted protease which cleaves host cell transcription factors essential for MHC class I and II antigen presentation. The identification of CPAF in this paper verifies the applicability of the described method for the identification of secreted proteins. We extend the findings by Zhong et al. by proteome studies of expression and turnover of C. trachomatis CPAF showing that the degradation of C. trachomatis D CPAF in the host cell is very limited. Furthermore, we show that two fragments of CPAF exist in C. pneumoniae as well as in C. trachomatis.

AB - Chlamydiae are obligate intracellular bacteria that are important human pathogens. The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified. Most likely, effector proteins are secreted in order to promote infection. Effector proteins cannot be identified by motif or similarity searches. As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of [35S]-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified Chlamydia. Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method. Two protein spots were identified among the candidates. These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D-PAGE profiles of whole lysates of infected cells but absent from purified Chlamydia. CPAF was recently identified by Zhong and colleagues as a secreted protease which cleaves host cell transcription factors essential for MHC class I and II antigen presentation. The identification of CPAF in this paper verifies the applicability of the described method for the identification of secreted proteins. We extend the findings by Zhong et al. by proteome studies of expression and turnover of C. trachomatis CPAF showing that the degradation of C. trachomatis D CPAF in the host cell is very limited. Furthermore, we show that two fragments of CPAF exist in C. pneumoniae as well as in C. trachomatis.

KW - Amino Acid Sequence

KW - Antibodies, Bacterial

KW - Base Sequence

KW - Cell Line

KW - Chlamydia trachomatis

KW - Chlamydophila pneumoniae

KW - Cloning, Molecular

KW - DNA, Bacterial

KW - Electrophoresis, Gel, Two-Dimensional

KW - Endopeptidases

KW - Hela Cells

KW - Humans

KW - Microscopy, Fluorescence

KW - Molecular Sequence Data

KW - Peptide Fragments

KW - Protease Inhibitors

KW - Species Specificity

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

M3 - Journal article

VL - 4

SP - 411

EP - 424

JO - Cellular Microbiology

JF - Cellular Microbiology

SN - 1462-5814

IS - 7

ER -

Shaw AC, Vandahl B, Larsen MR, Roepstorff P, Gevaert K, Vandekerckhove J et al. Characterization of a secreted Chlamydia protease. Cellular Microbiology. 2002;4(7):411-24.