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Open Access Research article

Modelling phagosomal lipid networks that regulate actin assembly

Mark Kühnel1, Luis S Mayorga12*, Thomas Dandekar3, Juilee Thakar4, Roland Schwarz5, Elsa Anes6, Gareth Griffiths1* and Jens Reich7

Author Affiliations

1 EMBL, Postfach 102209, 69117 Heidelberg, Germany

2 Laboratorio de Biología Celular y Molecular, IHEM-CONICET, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza, Argentina

3 Lehrstuhl für Bioinformatik, Biozentrum, Am Hubland, D-97074 Würzburg, Germany

4 Department of Physics, 104 Davey Laboratory, Pennsylvania State University, University Park, PA 16802, USA

5 Institut für Hygiene und Mikrobiologie, Josef-Schneider-Straße 2/Bau E1 97080 Würzburg, Germany

6 Molecular Pathogenesis Centre, Unit of Retrovirus and Associated Infections, Faculty of Pharmacy, University of Lisbon, Av. Forcas Armadas, 1600-083 Lisbon, Portugal

7 Max Delbrück Center, PO Box 74023810, D-13092 Berlin, Germany

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BMC Systems Biology 2008, 2:107  doi:10.1186/1752-0509-2-107

Published: 5 December 2008

Abstract

Background

When purified phagosomes are incubated in the presence of actin under appropriate conditions, microfilaments start growing from the membrane in a process that is affected by ATP and the lipid composition of the membrane. Isolated phagosomes are metabolically active organelles that contain enzymes and metabolites necessary for lipid interconversion. Hence, addition of ATP, lipids, and actin to the system alter the steady-state composition of the phagosomal membrane at the same time that the actin nucleation is initiated. Our aim was to model all these processes in parallel.

Results

We compiled detailed experimental data on the effects of different lipids and ATP on actin nucleation and we investigated experimentally lipid interconversion and ATP metabolism in phagosomes by using suitable radioactive compounds.

In a first step, a complex lipid network interconnected by chemical reactions catalyzed by known enzymes was modelled in COPASI (Complex Pathway Simulator). However, several lines of experimental evidence indicated that only the phosphatidylinositol branch of the network was active, an observation that dramatically reduced the number of parameters in the model. The results also indicated that a lipid network-independent ATP-consuming activity should be included in the model. When this activity was introduced, the set of differential equations satisfactorily reproduced the experimental data. On the other hand, a molecular mechanism connecting membrane lipids, ATP, and the actin nucleation process is still missing. We therefore adopted a phenomenological (black-box) approach to represent the empirical observations. We proposed that lipids and ATP influence the dynamic interconversion between active and inactive actin nucleation sites. With this simple model, all the experimental data were satisfactorily fitted with a single positive parameter per lipid and ATP.

Conclusion

By establishing an active 'dialogue' between an initial complex model and experimental observations, we could narrow the set of differential equations and parameters required to characterize the time-dependent changes of metabolites influencing actin nucleation on phagosomes. For this, the global model was dissected into three sub-models: ATP consumption, lipid interconversion, and nucleation of actin on phagosomal membranes. This scheme allowed us to describe this complex system with a relatively small set of differential equations and kinetic parameters that satisfactorily reproduced the experimental data.