Monitoring time-dependent maturation changes in purified phagosomes from Dictyostelium discoideum

Article Article scientifique Article Monitoring time-dependent maturation changes in purified phagosomes from Dictyostelium discoideum eng 2008 author Dieckmann Régis 500389 author Gopaldass Navin Andréw 634451 author Escalera Caroline 537681 author Soldati Thierry 0000-0002-2056-7931 135632 327-337 10.1007/978-1-59745-157-4_21 18425460 1064-3745 The amoeba Dictyostelium discoideum is an established model to study phagocytosis. The sequence of events leading to the internalization and degradation of a particle is conserved in D. discoideum compared to metazoan cells. As its small haploid genome has been sequenced, it is now amenable to genome-wide analysis including organelle proteomics. Therefore, we adapted to Dictyostelium the classical protocol to purify phagosomes formed by ingestion of latex beads particles. The pulse-chase protocol detailed here gives easy access to pure, intact, and synchronized phagosomes from representative stages of the entire process of phagosome maturation. Recently, this protocol was used to generate individual temporal profiles of proteins and lipids during phagosome maturation generating a proteomic fingerprint of six maturation stages (1). In addition, immunolabeling of phagosomes on a coverslip was developed to visualize and quantitate antigen distribution at the level of individual phagosomes. Animals Dictyostelium/metabolism Fluorescent Antibody Technique Phagocytosis/physiology Phagosomes/metabolism Time Factors Dewey 540 39 Département de biochimie 144 false Methods in molecular biology 445 7a598d0e-f141-406e-8dca-558582bfc30d Article (Published version) Public Dieckmann et al08.pdf 329671 application/pdf openAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie

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Monitoring time-dependent maturation changes in purified phagosomes from Dictyostelium discoideum	Methods in molecular biology			2008	635	431

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Escalera, Caroline