Study Guide: Endocytosis, Actin & Microfilaments Fall 2022

MCB 2210

Endocytosis

Endocytosis provides a route of entry for selected materials coming from outside the cell. These
materials may follow one of three distinct but closely related pathways. Bulk-phase endocytosis, or
pinocytosis (“cell drinking”), simply takes in bulk fluid from the surrounding medium, and phagocytosis
(“cell eating”) takes in large, insoluble aggregates of molecules, cell parts, or even whole cells. When it
comes to the third route, receptor-mediated endocytosis, substances taken in by this route are
recognized and bound to the cell surface by specific receptors. The receptors are usually glycoproteins
embedded in the plasma membrane, and molecules entering the cell by receptor-mediated endocytosis
become significantly more concentrated because they are tied to the cell surface and packed into
vesicles. Substances bind to their specific receptors and are then enclosed in pockets that invaginate
inward from the plasma membrane. The pockets pinch off from the plasma membrane as endocytic
vesicles, also called endosomes, and move into the cytoplasm. Once inside, the vesicles have various
fates depending on the type of substances enclosed within them. In some pathways, the endosomes
fuse with lysosomes, a process that exposes the proteins within them to hydrolysis by lysosomal
enzymes. Other pathways deliver endocytic vesicles to the Golgi complex, or on to other vesicles in the
cytoplasm.

Receptor-mediated endocytosis in mammalian cells has been studied experimentally in great detail.
Substances taken up by mammalian cells using this endocytic route include: growth factors, peptide
hormones, blood serum proteins, iron, vitamins and cholesterol. Please note - Pathogens and toxins,
including VIRUSES and the cholera and diphtheria toxins, ALSO use this pathway. Receptor-mediated
endocytosis is known to involve both microfilaments and microtubules. Pinching off pockets of the
plasma membrane and production of endosomes are dependent on microfilaments; the movement of
endosomes and their contents through the cytoplasm is powered by microtubules. For example,
microtubules lie in tracks extending in the directions followed by vesicles in their travels between the
Golgi complex and the plasma membrane. It has been repeatedly shown that vesicles can move in
both directions over the surface of the microtubules, with their movements powered by motor
proteins. As one would expect from these observations, vesicle movement ceases when cells are
exposed to the microtubule poison colchicine.

The Actin Cytoskeleton and Receptor-Mediated Endocytosis

The actin cytoskeleton is thought to play an important and active role in endocytosis for many reasons.
Endocytosis can be inhibited by drugs that block or inhibit the polymerization of actin. Actin and actin-
related proteins like ARP-3, cofilin, and others, are consistently localized to regions of the plasma
membrane undergoing endocytosis. The actin cytoskeleton has been implicated in calcium-dependent
vesicular traffic, and it is thought that actin polymerization processes play an important role during
vesicle budding and movement of endosomes through the cytoplasm. A number of labs have shown F-
actin-dependent movement of GFP-labeled early endosomes.
Actin & The Actin Cytoskeleton – the “Big Picture”

Here are some the “big picture” things you should know about actin and microfilaments (summary
taken from the recommended text book Essential Cell Biology, 4th Edition):

1. Actin filaments are helical polymers of globular actin monomers. They are more flexible than
microtubules and are generally found in bundles or networks.
2. Like microtubules, actin filaments are polarized, with a fast-growing plus end and a slow-
growing minus end. Assembly and disassembly are controlled by the hydrolysis of ATP tightly
bound to each actin monomer, and by various actin-binding and actin-related proteins
(thymosin, profilin, cofilin, etc.).
3. Actin bundling proteins control the behavior of actin filaments in vertebrate cells. Other proteins
cross-link actin filaments together in a gel-like meshwork within the cell cortex, the specialized
layer of actin-filament-rich cytoplasm just beneath the plasma membrane. Actin filaments can
also associate with myosin motor proteins to form contractile bundles, and they often form
tracks along which myosin motors transport organelles.
4. Cell crawling depends on cortical actin. Examples of actin-dependent cell crawling include
carnivorous amoebae searching for food, the advancing tip of a growing axon responding to
nerve growth factor, white blood cells migrating out of the blood into infected tissues to “search
and destroy” invading pathogens.

Behavior of actin during crawling: When the cell surface is pushed outward, the first step is
driven by actin polymerization. The leading edge of the cell extends thin, sheet-like lamellipodia,
which contain a dense network of actin filaments oriented so that most of the filaments have
their plus ends close to the plasma membrane. Many cells also extend thin, stiff protrusions
called filopodia, both at the leading edge and elsewhere on their surface. These are spikes of
about 0.1 µm wide and 5-10 µm long, each again oriented with plus ends of microfilaments
pointing outward. Both lamellipodia and filopodia are exploratory, motile structures that form
and retract with great speed, moving at 1 µm per second. They show the ability of actin
filaments to push out the plasma membrane without tearing it!