![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
|
|
|
|
|
||
A more recent version of this article appeared on October 1, 2002
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Submitted on April 18, 2002
Revised on July 2, 2002
Accepted on July 18, 2002
1 Department of Molecular Cell Research, Max-Planck-Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany
2 Department of Cell Physiology, Max-Planck-Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany
3 Laboratoire de Biochimie et Biophysique des Systèmes Intégrés, UMR5092-CNRS, CEA, 38054 Grenoble, France
4 Institute of Cell Biology, Ludwig-Maximilians University, Schillerstrasse 42, D-80336 Munich, Germany
5 Department of Molecular Cell Research, Max-Planck-Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany; and Department of Biological Sciences, Alexander Fleming Building, Imperial College of Science Technology and Medicine, Imperial College Road, London SW7 2AZ, UK
* Corresponding author. E-mail address: t.soldati{at}ic.ac.uk.
Molecular mechanisms of endocytosis in the genetically and biochemically tractable professional phagocyte Dictyostelium discoideum reveal a striking degree of similarity to higher eukaryotic cells. Pulse-chase feeding with LBs allowed purification of phagosomes at different maturation stages. Gentle ATP-stripping of an actin meshwork entrapping contaminating organelles resulted in a 10-fold increase in yield and purity, as confirmed by EM. Temporal profiling of signaling, cytoskeletal and trafficking proteins resulted in a complex molecular fingerprint of phagosome biogenesis and maturation. First, nascent phagosomes were associated with coronin, and rapidly received a lysosomal glycoprotein, LmpB. Second, at least two phases of delivery of lysosomal hydrolases (CatD and CP34) were accompanied by removal of plasma membrane components (PM4C4 and biotinylated surface proteins). Third, a phase of late maturation, preparing for final exocytosis of undigested material, included quantitative recycling of hydrolases and association with vacuolin. Also, lysosomal glycoproteins of the Lmp family showed distinct trafficking kinetics. The delivery and recycling of CatD was directly visualized by confocal microscopy. This heavy membrane traffic of cargos was precisely accompanied by regulatory proteins such as the Rab7 GTPases and the endosomal SNAREs Vti1 and VAMP7. This initial molecular description of phagocytosis demonstrates the feasibility of a comprehensive analysis of phagosomal lipids and proteins in genetically modified strains.
This article has been cited by other articles:
|
|
 |
|
  D. Gotthardt, V. Blancheteau, A. Bosserhoff, T. Ruppert, M. Delorenzi, and T. Soldati Proteomics Fingerprinting of Phagosome Maturation and Evidence for the Role of a G{alpha} during Uptake Mol. Cell. Proteomics, December 1, 2006; 5(12): 2228 - 2243. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Jacobs, L. V. DeSouza, H. Samaranayake, R. E. Pearlman, K. W. M. Siu, and L. A. Klobutcher The Tetrahymena thermophila Phagosome Proteome Eukaryot. Cell, December 1, 2006; 5(12): 1990 - 2000. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Hatsuzawa, T. Tamura, H. Hashimoto, H. Hashimoto, S. Yokoya, M. Miura, H. Nagaya, and I. Wada Involvement of Syntaxin 18, an Endoplasmic Reticulum (ER)-localized SNARE Protein, in ER-mediated Phagocytosis Mol. Biol. Cell, September 1, 2006; 17(9): 3964 - 3977. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Touret, P. Paroutis, and S. Grinstein The nature of the phagosomal membrane: endoplasmic reticulum versus plasmalemma J. Leukoc. Biol., June 1, 2005; 77(6): 878 - 885. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. L. Lee, M.-K. Kim, A. D. Schreiber, and S. Grinstein Role of Ubiquitin and Proteasomes in Phagosome Maturation Mol. Biol. Cell, April 1, 2005; 16(4): 2077 - 2090. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Bharathi, S. K. Pallavi, R. Bajpai, B. S. Emerald, and L. S. Shashidhara Genetic characterization of the Drosophila homologue of coronin J. Cell Sci., April 15, 2004; 117(10): 1911 - 1922. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Lefkir, M. Malbouyres, D. Gotthardt, A. Ozinsky, S. Cornillon, F. Bruckert, A. A. Aderem, T. Soldati, P. Cosson, and F. Letourneur Involvement of the AP-1 Adaptor Complex in Early Steps of Phagocytosis and Macropinocytosis Mol. Biol. Cell, February 1, 2004; 15(2): 861 - 869. [Abstract] [Full Text] [PDF]
|
|
