The budding of a vesicle from its parent membrane is driven by the polymerization of soluble protein complexes on the membrane to form a proteinaceous vesicle coat (Figure 1a). Interactions between the cytosolic portions of integral mem brane proteins and the vesicle coat gather the appropriate cargo proteins into the forming vesicle. Thus the coat gives curvature to the membrane to form a vesicle and acts as the filter to determine which proteins are admitted into the vesicle.
Fig1. Overview of vesicle budding and fusion with a target membrane. (a) Budding is initiated by recruitment of a small GTP-binding protein to a patch of donor membrane. Complexes of coat proteins in the cytosol then bind to the cytosolic domain of membrane cargo proteins, some of which also act as receptors that bind soluble proteins in the lumen, thereby recruiting luminal cargo proteins into the budding vesicle. (b) After being released and shedding its coat, a vesicle fuses with its target membrane in a process that involves inter action of cognate SNARE proteins.
Proteins responsible for the eventual fusion of a vesicle with the target membrane, known as v-SNAREs, are incorporated into the vesicle membrane during assembly of the vesicle coat. After the coat is shed from a completed vesicle, v-SNARE proteins embedded in the vesicle membrane become accessible to join with cognate t-SNAREs in the target membrane to which the vesicle is docked. This joining brings the membranes into close apposition, allowing the two bi layers to fuse (Figure 1b). Regardless of target organelle, all transport vesicles use v-SNAREs and t-SNAREs to fuse.
Three major types of coated vesicles have been characterized, each with a different type of protein coat and each formed by reversible polymerization of a distinct set of protein subunits (Table 14-1). Each type of vesicle, named for its primary coat proteins, transports cargo proteins from particular parent organelles to particular target organelles:
● COPII vesicles transport proteins from the ER to the Golgi.
● COPI vesicles mainly transport proteins in the retrograde direction between Golgi cisternae and from the cis-Golgi back to the ER.
● Clathrin-coated vesicles transport proteins from the plasma membrane (cell surface) and the trans-Golgi network to late endosomes.
Table1. Coated Vesicles Involved in Protein Trafficking
Every vesicle-mediated trafficking step is thought to use some kind of vesicle coat; however, a specific coat protein complex has not been identified for every type of vesicle. For example, vesicles that move proteins from the trans-Golgi to the plasma membrane during either constitutive or regulated secretion exhibit a uniform size and morphology, which suggests that their formation is driven by assembly of a regular coat structure, yet researchers have not identified specific coat proteins surrounding these vesicles.
The general scheme of vesicle budding shown in Figure 1a applies to all three known types of coated vesicles. Experiments with isolated or artificial membranes and purified coat proteins have shown that polymerization of the coat proteins on the cytosolic face of the parent membrane is necessary to produce the high curvature of the membrane that is typical of a transport vesicle about 50 nm in diameter. Electron micrographs of in vitro budding reactions often reveal structures that exhibit discrete regions of the parent membrane bearing a dense coat accompanied by the curvature characteristic of a completed vesicle (Figure 2). Such structures, usually called vesicle buds, appear to be intermediates that are visible after the coat has begun to polymerize but before the completed vesicle pinches off from the parent membrane. The polymerized coat proteins are thought to form a curved lattice that drives the formation of a vesicle bud by adhering to the cytosolic face of the membrane.
Fig2. Vesicle buds can be visualized during in vitro budding reactions. When purified COPII coat components are incubated with isolated ER vesicles or artificial phospholipid vesicles (liposomes), polymerization of the coat proteins on the vesicle surface induces emergence of highly curved buds. In this electron micrograph of an in vitro budding reaction, note the distinct mem brane coat, visible as a dark protein layer, present on the vesicle buds. [Republished with permission of Elsevier, from Matsuoka, K. et al., “COPII-coated vesicle formation reconstituted with purified coat proteins and chemically defined liposomes,” 1998, Cell 93(2):263–275; permission conveyed through the Copyright Clearance Center, Inc.]
