 | |  | |  | |  | |  | |  | |  | |  |
| IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711 |
Int J Clin Exp Med 2(3):233-247;2009.
Original Article
A systems biology approach to study the phagosomal proteome modulated by
mycobacterial infections
Prahlad K. Rao, Christoher R. Singh, Chinnaswamy Jagannath, and Qingbo Li
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, Chicago, IL 60607, USA; Department of Pathology and
Laboratory Medicine, University of Texas Health Sciences Center, Houston, TX 77030, USA; Department of Microbiology and
Immunology, University of Illinois at Chicago, Chicago, IL 60612, USA.
Received July 1, 2009; accepted September 8, 2009; available online September 31, 2009
Abstract: Systems biology and proteomics have recently contributed significantly to the insight into the biogenesis and
immunity-related functions of the phagosome. To gain insight into the modulation of the phagosomal proteome by the wild-type
Mycobacterium tuberculosis H37Rv reference strain, an attenuated mutant of the H37Rv strain, and the BCG Pasteur vaccine strain, we
employed the nano-liquid chromatography/LTQ-FTMS based proteomics approach and a systems biology approach to analyze the
bacillus-containing phagosomes purified from the bone-marrow-derived BMA3.A3 macrophages infected with the three different
mycobacterial strains. We identified 322 proteins at a false-discovery rate of 2%. These proteins were quantified with a label-free
proteomics method. All but one of these proteins is mouse proteins. The gene ontology analysis of these mouse proteins suggests
that lysosomal proteins represented <3% of the detected proteins, supporting the observation that these mycobacterial strains inhibit or
limit the phagosome maturation process. The results also indicate that the endop-lasmic reticulum (ER) proteins do not constitute a
major part of the phagosome proteome, supporting the phago-some maturation model of the role of ER in phagosome biogenesis.
This phagosome maturation model is in contrast to the phagocytosis model which predicts that half of the phagosome membrane is
derived from ER. This pilot study demonstrates that a combination of proteomics, multivariate analysis, and systems biology promises
to bring forward new insights into the mycobacterial pathogenesis and the interconnected phagosome biology. (IJCEM907001).
Key Words: Phagosome, systems biology, mycobacterium tuberculosis, proteomics, macrophage
Full Text PDF
Address all correspondence to:
Qingbo Li, PhD,
Department of Microbiology and Immunology
University of Illinois at Chicago
Chicago, IL 60612
USA
Tel: 312-413-9301; Fax: 312-413-9303
E-Mail: qkli@uic.edu
Original Article
A systems biology approach to study the phagosomal proteome modulated by
mycobacterial infections
Prahlad K. Rao, Christoher R. Singh, Chinnaswamy Jagannath, and Qingbo Li
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, Chicago, IL 60607, USA; Department of Pathology and
Laboratory Medicine, University of Texas Health Sciences Center, Houston, TX 77030, USA; Department of Microbiology and
Immunology, University of Illinois at Chicago, Chicago, IL 60612, USA.
Received July 1, 2009; accepted September 8, 2009; available online September 31, 2009
Abstract: Systems biology and proteomics have recently contributed significantly to the insight into the biogenesis and
immunity-related functions of the phagosome. To gain insight into the modulation of the phagosomal proteome by the wild-type
Mycobacterium tuberculosis H37Rv reference strain, an attenuated mutant of the H37Rv strain, and the BCG Pasteur vaccine strain, we
employed the nano-liquid chromatography/LTQ-FTMS based proteomics approach and a systems biology approach to analyze the
bacillus-containing phagosomes purified from the bone-marrow-derived BMA3.A3 macrophages infected with the three different
mycobacterial strains. We identified 322 proteins at a false-discovery rate of 2%. These proteins were quantified with a label-free
proteomics method. All but one of these proteins is mouse proteins. The gene ontology analysis of these mouse proteins suggests
that lysosomal proteins represented <3% of the detected proteins, supporting the observation that these mycobacterial strains inhibit or
limit the phagosome maturation process. The results also indicate that the endop-lasmic reticulum (ER) proteins do not constitute a
major part of the phagosome proteome, supporting the phago-some maturation model of the role of ER in phagosome biogenesis.
This phagosome maturation model is in contrast to the phagocytosis model which predicts that half of the phagosome membrane is
derived from ER. This pilot study demonstrates that a combination of proteomics, multivariate analysis, and systems biology promises
to bring forward new insights into the mycobacterial pathogenesis and the interconnected phagosome biology. (IJCEM907001).
Key Words: Phagosome, systems biology, mycobacterium tuberculosis, proteomics, macrophage
Full Text PDF
Address all correspondence to:
Qingbo Li, PhD,
Department of Microbiology and Immunology
University of Illinois at Chicago
Chicago, IL 60612
USA
Tel: 312-413-9301; Fax: 312-413-9303
E-Mail: qkli@uic.edu
