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  • Review Article
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Shaping cups into phagosomes and macropinosomes

Nature Reviews Molecular Cell Biology volume 9, pages 639–649 (2008)Cite this article

Key Points

  • Phagocytosis is the ingestion by cells of particles or other cells. Receptors in membranes of phagocytic cells organize the advance of the plasma membrane and the actin cytoskeleton around target particles, forming intracellular membrane-bounded compartments called phagosomes.

  • Macropinocytosis is the ingestion by cells of extracellular solutes and fluid into 0.2–10-μm diameter vesicles, or macropinosomes. Macropinosomes can form spontaneously or in response to activation of cell-surface receptors.

  • Common signalling mechanisms organize the construction of phagocytic or macropinocytic cup-shaped invaginations of the plasma membrane. Receptors, GTPases of the Ras superfamily and membrane phospholipids regulate the component activities of actin-filament assembly, disassembly and contraction, as well as the fusion of membrane vesicles with cup membranes.

  • Imaging of molecular dynamics in living cells indicates that phagocytic and macropinocytic cups exhibit distinct patterns of signalling that correspond to the early and late stages of their formation.

  • Activated receptor complexes (short-range signals) generate diffusible signal molecules (medium-range signals) that are subject to feedback regulation and that, at suprathreshold concentrations, can activate transitions from early to late stages of signalling. Phospholipids generated in the confines of a forming cup integrate and amplify signalling in that region of the membrane.

  • Receptor-mediated signal transduction for phagocytosis and macropinocytosis is modulated by cell structure and is conditional on feedback regulation that is related to cup integrity, particle stiffness and particle shape.

Abstract

The ingestion of particles or cells by phagocytosis and of fluids by macropinocytosis requires the formation of large endocytic vacuolar compartments inside cells by the organized movements of membranes and the actin cytoskeleton. Fc-receptor-mediated phagocytosis is guided by the zipper-like progression of local, receptor-initiated responses that conform to particle geometry. By contrast, macropinosomes and some phagosomes form with little or no guidance from receptors. The common organizing structure is a cup-shaped invagination of the plasma membrane that becomes the phagosome or macropinosome. Recent studies, focusing on the physical properties of forming cups, indicate that a feedback mechanism regulates the signal transduction of phagocytosis and macropinocytosis.

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Figure 1: The movements of phagocytosis and macropinocytosis.
Figure 2: Short- and medium-range signalling by activated Fc and EGFR.
Figure 3: Distinct patterns of signalling in phagocytic cups.
Figure 4: Context-dependent signal transduction in cups.

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Acknowledgements

I am grateful for the suggestions of S. Yoshida and A. Hoppe, and for funding from the National Institute of Allergy and Infectious Disease (NIAID) and the National Institutes of Health (NIH).

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  1. Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, 48109-5620, Michigan, USA

    Joel A. Swanson

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Supplementary information S1 (movie) | Activation of Rac1 during M–CSF-induced macropinocytosis.

Bone marrow-derived mouse macrophages expressing fluorescence resonance energy transfer (FRET) probes for Rac1 activity (yellow fluorescent protein (YFP)–Rac1 and cyan fluorescent protein (CFP)–PBD (p21– binding domain)) were stimulated with 200 ng/ml macrophage colony-stimulating factor (M–CSF) just prior to the first frame of the video sequence. The left panel shows time-lapse, phase-contrast microscopy immediately following the addition of M–CSF. Extensive ruffling is evident as the dark folds of plasma membrane that form and migrate from the cell periphery. The right panel shows the same phase–contrast sequence with an overlay showing GTP–bound Rac1, as detected by FRET microscopy. Rac1 was activated initially throughout the periphery of the cell, but its activation soon became limited to the forming macropinosomes. Images were collected every 20 sec and played back at 6 frames/sec (that is, 100x real time). Sequence courtesy of S. Yoshida, University of Michigan Medical School, Ann Arbor, Michigan, USA. (MOV 237 kb)

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Glossary

Clathrin

A protein that facilitates endocytosis of receptors in small (>0.1 μm in diameter) vesicles by forming and reorganizing a coat on the cytoplasmic face of a membrane.

Fc receptor

A class of cell-surface receptors that bind to the Fc domain of immunoglobulin molecules such as immunoglobulin G.

Opsonize

The coating of a particle with molecules that renders it capable of being bound and ingested by phagocytic cells. From Greek, meaning 'to prepare a meal'.

Recycling endosomes

A subclass of endocytic vesicles that communicate by vesicle fusion with other endocytic compartments and regions of the plasma membrane, including phagocytic cups.

Ruffle

A thin, sheet-like protrusion of the plasma membrane that extends from the cell surface through the formation and growth of a branched network of actin filaments. Ruffles either retract into the cytoplasm or close into macropinosomes.

Gelation

The formation of a crosslinked matrix of polymer. In the actin cytoskeleton, actin filaments are bridged by crosslinking proteins into a gel. The regulated dissolution of that matrix (solation) can be coupled with the activation of contractile proteins to effect motility.

Lamellipodium

A sheet-like protrusion of the cell surface that contains a branched network of actin filaments that extends along surfaces during cell motility. Lamellipodia are structurally analogous to ruffles and phagocytic cups.

Microdomain

A small region of membrane, rich in cholesterol or sphingolipids, to which certain classes of lipid-anchored peripheral membrane proteins, such as Src-family kinases, localize preferentially.

Pleckstrin-homology domain

A structural domain that is common in many signalling proteins and that can bind specific phosphoinositol sugars, including those that comprise membrane phosphoinositides.

Ras superfamily

A large group of small proteins that share structural features and the capabilities of regulated binding to GTP, GDP, membranes, proteins that regulate the species of bound nucleotide and various additional effector proteins.

Multivesicular body

An intracellular membranous compartment that contains intracellular vesicles that are derived from its delimiting membrane. Multivesicular bodies communicate by vesicle fusion with endosomes, lysosomes and sometimes also with the plasma membrane.

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Swanson, J. Shaping cups into phagosomes and macropinosomes. Nat Rev Mol Cell Biol 9, 639–649 (2008). https://doi.org/10.1038/nrm2447

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