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NMF/cellbio/cytomemb
I. Introduction
Membrane divides the cytoplasm of eukaryotic cells into distinct compartments. Membrane-bound structures (organelles) are found in all eukaryotic cells.
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Endocytic pathways move materials into cells. Sorting signals are recognized by receptors and target proteins to specific sites.
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Autoradiography (pulse-chase experiment; use of labelled amino acids). Biochemical analyses of subcellular fractions (microsomes of endoplasmic reticulum (ER) and Golgi complex (GC). Study of genetic mutants.
NMF/cellbio/cytomemb
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Synthesis of steroids in endocrine cells. Detoxification of organic compounds in liver cells. Release of glucose from glucose-6-phosphate in liver cells. Sequestration of Ca2+.
NMF/cellbio/cytomemb
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Rough ER (RER). Network of flattened cisternae with ribosomes on the cytosolic surface. The polarity of organelles in some secretory cells reflects the flow from the site of synthesis to the site of discharge. Proteins synthesized on ribosomes of RER include secretory proteins, integral membrane proteins and proteins of organelles. Proteins synthesized on free ribosomes include cytosolic proteins, peripheral membrane proteins, nuclear proteins and proteins found in mitochondria and chloroplasts.
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Membrane biosynthesis in the ER: Lipids are inserted into existing membranes. Asymmetry of proteins and lipids is maintained during membrane trafficking. Some integral membrane proteins are synthesized on ribosome of the ER and translocated into the ER membrane cotranslationally. Integral membrane proteins have hydrophobic stop-transfer sequences that cause them to bind to the inner surface of the translocon.
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Synthesis of membrane lipids: Lipids inserted into the outer leaflet of the ER are modified by: Conversion of one type of phospholipid to another type. Selective inclusion in nascent vesicles. Phospholipid transfer proteins that move lipids between membranes.
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Glycosylation in the RER Carbohydrates are added by glycosyltransferases to almost all proteins produced on membrane-bound ribosomes. Sugars are attached in an ordered sequence. The core carbohydrate chain is assembled on a lipid carrier, dolichol phosphate. The core carbohydrate is transferred to the polypeptide by oligosaccharyltransferase and is modified en route to the Golgi complex.
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The first step in vesicular transport: 1. Membrane and luminal proteins move to the tips of RER. 2. Transitional elements i.e. RER tips form the first transport vesicles.
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Stacks of disc-shaped flattened cisternae. The cis face of the Golgi faces the ER; the trans face is on the opposite end of the stack. The Golgi networks are processing and sorting stations where proteins are modified, segregated and then shipped in different directions. The cis Golgi network (CGN) sorts proteins for the ER or the next Golgi station. The trans Golgi network (TGN) sorts proteins to various cellular destinations.
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Glycosylation in the Golgi complex: Synthesis of complex polysaccharides takes place in the Golgi. Oligosaccharides attach to glycoproteins and glycolipids in the Golgi. Vesicular transport within the Golgi complex: two views. In the maturation model, cisternae formed on the cis face are transient and physically move to the trans face Alternative model: material moves through permanent Golgi stacks in vesicles.
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VI.
Materials are carried between compartments by various protein-coated vesicles. Three major types of protein-coated vesicles: 1. COPII-coated vesicles move materials from the ER to the Golgi. 2. COPI-coated vesicles move materials from the Golgi to the ER. 3. Clathrin-coated vesicles move materials from the trans Golgi to endosomes, lysosomes and plant vacuoles, also materials from plasma membrane to organelles.
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Exocytosis The terminal stage of secretion. Cells engaged in extensive secretion have numerous secretory vesicles at the apical side. The process of exocytosis is triggered by local increases in Ca2+ concentration. Fusion proteins found on the vesicle and membrane mediate the formation of fusion pores. Vesicles may or may not become absorbed into the plasma membrane.
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VII. Lysosomes
Lysosomes contain hydrolases that can digest every kind of biological molecule. Internal proton concentration kept high by H+ATPases (optimum actv. at pH 4.6). Glycosylated proteins, Igp-A and Igp-B may protect the lysosome from self-digestion. Lysosomes are identified by the presence of the enzyme acid phosphatase. Lysosomes are involved in two major cell functions: phagocytosis (macrophage and neutrophiles) and autophagy.
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Primary lysosome fuse with either phagocytic or autophagic vesicles, forming residual bodies that undergo excocytosis or are retained in the cell as lipofuscin granules.
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IX. Proteasomes
Proteasomes are multiprotein complexes that rid the cell of abnormal proteins and degrade individual cell proteins that are no longer needed. Found in nucleus and cytoplasm Proteins to be degraded are marked by linkage to ubiquitin.
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Plant cells contain large vacuoles (~90% of cell volume) for the storage of compounds Transport systems in the tonoplast (vacuole membrane) accumulate material in the vacuole, attracting water by osmosis; this in turn generates turgor pressure (support for the cell) Plant vacuoles, like lysosome of animal cells (end point for biosynthetic pathways & may contain acid hydrolases)
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