Endocytosis membrane components

The process by which cells take up large molecules is called endocytosis. Some of these molecules (eg, polysaccharides, proteins, and polynucleotides), when hydrolyzed inside the cell, yield nutrients. Endocytosis provides a mechanism for regulating the content of certain membrane components, hormone receptors being a case in point. Endocytosis can be used to learn more about how cells function. DNA from one cell type can be used to transfect a different cell and alter the latter s function or phenotype. A specific gene is often employed in these experiments, and this provides a unique way to smdy and analyze the regulation of that gene. DNA transfection depends upon endocytosis endocy-... 

Retrieval of membrane components in the secretory pathway through receptor-mediated endocytosis (RME) isaclathrin-coat-dependent process 155... 

Retrieval of membrane components in the secretory pathway through receptor-mediated endocytosis (RME) is a clathrin-coat-dependent process [5]. The clathrin coat provides stability to the vesicle core and allows uptake of specific membrane proteins for reuse or degradation. RME shows a remarkable degree of specificity, allowing cells to internalize with astonishing efficiency only those selected molecules independent of their extracellular concentration. 

The problem we have not yet touched upon is how components can specifically move from one cellular component to another. Both the entry and the exit of SFV spike proteins are dependent on a number of such cellular processes. The newly synthesized spike proteins move from the ER to the Golgi complex and then to the cell surface. The cell surface membrane is continuously retrieved by endocytosis into endosomes. From here the endocytosed membrane components probably recycle back to the cell surface, but some components may also be channeled into lysosomes for degradation. Especially in cells with secretory activity, the recycling pathway from the cell surface also includes the Golgi complex (see Farquhar and Palade, 1981). 

According to the classical view, synaptic vesicles completely flatten during exo-cytosis, which is followed by retrieval of the membrane components by clathrin-dependent endocytosis. Evidence for direct retrieval (kiss-and-run) was provided... 

Figure 7.7-2. Liposome-cell interactions. Drug-loaded liposomes can specifically (A) or nonspecificaiiy (B) adsorb onto the ceil surface. Liposomes can also fuse with the cell membrane (C) and release their contents into the cell cytoplasm, or they can be destabilized by certain cell membrane components when adsorbed on the surface (D) so that the released drug can enter the cell via micro-pinocytosis. Liposome can undergo the direct or transfer-protein mediated exchange of lipid components with the cell membrane (E) or be taken up by specific or nonspecific endocytosis (F). In the case of endocytosis, a liposome can be delivered by the endosome into the lysosome (G), or en route to the lysosome, the liposome can provoke endosome destabilization (H), which results in drug liberation into the cytoplasm. (With permission from Ref. 29.)...

A typical feature of caveola-mediated endocytosis is the formation of non-coated invaginations composed of detergent-resistant membrane components rich in cholesterol and sphingolipids, known as lipid rafts [56]. The importance of the caveola-mediated mechanism in the PTD-mediated internalization was confirmed in an experiment where the cellular uptake of Tat peptide was affected by drugs that either disrupt lipid rafts or alter caveolar trafficking [57]. Moreover, Tat-PTD-fused protein showed colocalization with a marker of caveolar uptake, caveolin, further strengthening the importance of the mechanism in the PTD-mediated internalization [57]. 

Biochemical characterization of clathrin-coated vesicles revealed that their major coat components are clathrin and various types of adaptor complexes. Clathrin assembles in triskelions that consist of three heavy chains of approximately 190 kDa and three light chains of 30 40 kDa. Four types of adaptor complexes have been identified to date, AP-1, AP-2, AP-3 and AP-4 (AP for adaptor protein). Whereas AP-1, AP-3 and AP-4 mediate sorting events at the TGN and/or endosomes, AP-2 is involved in endocytosis at the plasma membrane. Each adaptor complex is a hetero-tetrameric protein complex, and the term adaptin was extended to all subunits of these complexes. One complex is composed of two large adaptins (one each of y/a/S/s and [31-4, respectively, 90-130 kDa), one medium adaptin (pi -4, <50 kDa), and one small adaptin (ol-4, <20 kDa). In contrast to AP-1, AP-2 and AP-3, which interact directly with clathrin and are part of the clathrin-coated vesicles, AP-4 seems to be involved in budding of a certain type of non-clathrin-coated vesicles at the TGN. 

Membranes and their components are dynamic structures. The lipids and proteins in membranes undergo turnover there just as they do in other compartments of the cell. Different lipids have different turnover rates, and the turnover rates of individual species of membrane proteins may vary widely. The membrane itself can turn over even more rapidly than any of its constituents. This is discussed in more detail in the section on endocytosis. 

Most cells release macromolecules to the exterior by exocytosis. This process is also involved in membrane remodeling, when the components synthesized in the Colgi apparatus are carried in vesicles to the plasma membrane. The signal for exocytosis is often a hormone which, when it binds to a cell-surface receptor, induces a local and transient change in Ca concentration. Ca triggers exocytosis. Figure 41—16 provides a comparison of the mechanisms of exocytosis and endocytosis. 

There are receptors (TfRs) on the surfaces of many cells for transferrin, it binds to these receptors and is internalized by receptor-mediated endocytosis (compare the fate of LDL Chapter 25). The acid pH inside the lysosome causes the iron to dissociate from the protein. The dissociated iron leaves the endosome via DMTl to enter the cytoplasm. Unlike the protein component of LDL, apoTf is not degraded within the lysosome. Instead, it remains associated with its receptor, returns to the plasma membrane, dissociates from its receptor, reenters the plasma, picks up more iron, and again delivers the iron to needy ceils. 

Fig. 1 Bioresponsive polyplexes. (a) Systemic circulation of shielded polyplexes in blood stream and attachment to cell surface receptor (b) endocytosis into endosomes, deshielding by cleavage of PEG linkers and activation of membrane-destabilizing component by acidic pH or other means (c) endosomal escape into cytosol (d) siRNA transfer to form a cytosolic RNA-induced silencing complex complex (e) cytosolic migration and intranuclear import of pDNA (/) presentation of pDNA in accessible form to the transcription machinery...

Infection of CD4+ cells commences via interaction between gp 120 and the CD4 glycoprotein, which effectively acts as the viral receptor. Entry of the virus into the cell, which appears to require some additional cellular components, occurs via endocytosis and/or fusion of the viral and cellular membranes. The gp 41 transmembrane protein plays an essential role in this process. 

Under some circumstances, lysosomal hydrolases may fail to be properly packaged in the TGN, so they enter the default pathway to the cell surface, where they are secreted. Although these hydrolases do little harm at the nearly neutral pH of most extracellular fluids, they can also be returned to lysosomes by a pathway known as receptor-mediated endocytosis. In this pathway, M6P receptors are sent to the plasma membrane, where they bind escaped lysosomal hydrolases and bring them back to lysosomes through the early and late endosomes. Receptor-mediated endocytosis is a major component of the endocytic pathways for trafficking of membrane proteins and merit more detailed consideration. 

As noted above, synaptic vesicles are not typically generated at the level of the TGN. Instead, they are assembled from endocytosed material retrieved from the synaptic plasma membrane. Synaptic vesicle and plasma membrane lipids and proteins are synthesized in the endoplasmic reticulum and modified in the Golgi apparatus, where they are then packaged in secretory vesicles. These synaptic precursors are delivered to the plasma membrane from the cell body by the constitutive secretory pathway. Synaptic vesicle proteins must be retrieved by clathrin-mediated synaptic vesicle endocytosis, a variant of RME with some neuron-specific components. Once the vesicle sheds its clathrin coat, the uncoated vesicle fuses with a... 

However, an alternative pathway that bypasses clathrin-mediated endocytosis and EEs appears to be available as well. This model of endocytosis known as kiss and run or its variant kiss and stay have attracted increasing interest in recent years [74] (Fig. 9-9B). Kiss and run has been directly demonstrated with dense-core granules in neuroendocrine cells [84, 85], and this model would explain some observations that are not readily accommodated by the classical pathway. The kiss and run model proposes that neurotransmitters are released by a transient fusion pore, rather than by a complete fusion with integration of the synaptic vesicle components into the plasma membrane. Synaptic membrane proteins never lose their association and the vesicle reforms when the pore closes. As a result, the empty vesicle can be refilled and reused without going through clathrin-mediated endocytosis and sorting in the EEs. 

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