High-resolution dissection of phagosome maturation reveals distinct membrane trafficking phases

Autor: Franz Bruckert, Thierry Soldati, Hans-Jörg Warnatz, Oliver Henschel, Daniel Gotthardt, Michael Schleicher
Jazyk: angličtina
Rok vydání: 2002
Předmět:
Thiazoles/metabolism
Adenosine Triphosphate/metabolism
Coronin
Vesicular Transport Proteins
Carrier Proteins/metabolism
Cathepsin D
R-SNARE Proteins
Rab GTP-Binding Proteins/metabolism
Adenosine Triphosphate
Phagosomes
Dictyostelium
Dictyostelium/cytology/metabolism
Cytoskeleton
Phagosome
Microfilament Proteins
Cell Membrane/chemistry/metabolism
Qb-SNARE Proteins
Lipids
Endocytosis
Cell biology
Biochemistry
Fluorescent Dyes/metabolism
Thiazolidines
Biological Markers
Endocytosis/physiology
SNARE Proteins
Endosome
Microfilament Proteins/metabolism
Lipids/chemistry
Biology
Karyopherins
Exocytosis
Article
Phagosome maturation
Animals
Molecular Biology
Biological Transport/physiology
Fluorescent Dyes
Phagosomes/chemistry/metabolism/ultrastructure
Proteins/chemistry/metabolism
Cell Membrane
Membrane Proteins
Proteins
rab7 GTP-Binding Proteins
Biological Transport
Cell Biology
Cathepsin D/metabolism
Bridged Bicyclo Compounds
Heterocyclic
Lipid Metabolism
Thiazoles
Bicyclo Compounds
Heterocyclic/metabolism
Membrane protein
rab GTP-Binding Proteins
Karyopherins/metabolism
biology.protein
Membrane Proteins/metabolism
Carrier Proteins
Biomarkers
Zdroj: Molecular Biology of the Cell : the Official Publication of the American Society for Cell Biology
Molecular Biology of the Cell, Vol. 13, No 10 (2002) pp. 3508-20
ISSN: 1059-1524
Popis: 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 latex beads allowed purification of phagosomes at different stages of maturation. Gentle ATP stripping of an actin meshwork entrapping contaminating organelles resulted in a 10-fold increase in yield and purity, as confirmed by electron microscopy. 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 (cathepsin D [CatD] and cysteine protease [CPp34]) 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.
Databáze: OpenAIRE
Externí odkaz: