Endoplasmic reticulum membrane-associated synthesis

The pathways diverge at phosphatidate. In the synthesis of triacylglycerols, phosphatidate is hydrolyzed by a specific phosphatase to give a diacylglycerol (DAG). This intermediate is acylated to a triacylglycerol in a reaction that is catalyzed by diglyceride acyltransferase. Both enzymes are associated in a triacylglycerol synthetase complex that is bound to the endoplasmic reticulum membrane. 

The biosynthesis in yeast of two enzymes that are D-mannoproteins has been studied. A membrane-associated isozyme of invertase (EC 3.2.1.26) has been shown to be a precursor of the external invertase.190 Its molecular weight, as determined by SDS-poly(acrylamide) gel electrophoresis, is 50,000, that is, smaller than that of the external invertase, and it correlates well with the presence of only the inner-core sugars of the final form. It is strictly bound to membranes, possibly those of the endoplasmic reticulum, and it can be completely split191 by endo-/3-N-acetylglucosaminidase H (EC 3.2.1.30). The addition of tunicamycin, which specifically inhibits formation of d-GIcNAc-PP-DoI, inhibits synthesis of external invertase, as well as further formation of the membrane-associated form, which completely disappears after addition of the antibiotic.190 In these aspects, the synthesis of this extracellular enzyme follows the pathway for secreted glycoproteins in animal systems. 

Endoplasmic reticulum. A system of double membranes in the cytoplasm that is involved in the synthesis of transported proteins. The rough endoplasmic reticulum has ribosomes associated with it. The smooth endoplasmic reticulum does not. 

Milgrom and co-workers [105] recently developed an immunogold method for detection of PR in the rabbit uterus and have examined the effect of hormone addition on receptor localization at the ultrastructural level. PR were found to be predominantly nuclear in the presence and absence of hormone, but a small amount was detectable in the cytoplasm which was not apparent at the light microscopical level. These cytoplasmic PR were localized over endoplasmic reticulum and clusters of free ribosomes and may likely represent newly synthesized protein. No PR were located in the plasma membrane. Within the nucleus, unoccupied PR were associated with condensed chromatin which became more dispersed after hormone addition. These ultrastructural studies indicate that steroid-free PR translocate from their site of synthesis in the cytoplasm to the nucleus in a hormone independent manner, and that addition of hormone changes their intranuclear localization. 

CYP enzymes are a large and diverse superfamily of haemoproteins. Primarily membrane-associated proteins, they are located in the inner membrane of mitochondria and the endoplasmic reticulum of cells, and metabolise thousands of endogenous and exogenous compounds. Most of these enzymes can metabolise multiple substrates, and many can catalyse multiple reactions. While prevalent in the liver, CYP enzymes are also present in most other tissues of the body, and play an important role in hormone synthesis and breakdown (including oestrogen and testosterone synthesis and metabolism), cholesterol synthesis and vitamin D metabolism. Hepatic CYPs are the most widely smdied. 

An extensive array of internal membranes in eukaryotes creates compartments within a cell for distinct biochemical functions. For instance, a double membrane surrounds the nucleus, the location of most of the cell s genetic material, and the mitochondria, the location of most ATP synthesis. A single membrane defines the other internal compartments, such as the endoplasmic reticulum. Some compartments can exchange material by the process of membrane budding and fusion. As with all membranes, the proteins associated with these membranes determine the specific biochemical function. Specific amino acid sequences in the proteins direct these molecules to the appropriate compartment. 

The answer is e. (Murray, pp 452-467. Scriver, pp 3-45. Sack, pp 1—40. Wilson, pp 101-120.) Protein synthesis occurs in the cytoplasm, on groups of free ribosomes called polysomes, and on ribosomes associated with membranes, termed the rough endoplasmic reticulum. However, proteins destined for secretion are only synthesized on ribosomes of the endoplasmic reticulum and are synthesized in such a manner that they end up inside the lumen of the endoplasmic reticulum. From there the secretory proteins are packaged in vesicles. The Golgi apparatus is involved in the glycosylation and packaging of macromolecules into membranes for secretion. 

The answer is c. (Murray, pp 452—467. Scriver, pp 3—45. Sack, pp 1—40. Wilson, pp 101-120.) Prokaryotic ribosomes have a sedimentation coefficient of 70S and are composed of SOS and 30S subunits. Eukaryotic cytoplasmic ribosomes, either free or bound to the endoplasmic reticulum, are larger—60S and 40S subunits that associate to an SOS ribosome. Nuclear ribosomes are attached to the endoplasmic reticulum of the nuclear membrane. Ribosomes in chloroplasts and mitochondria of eukaryotic cells are more similar to prokaryotic ribosomes than to eukaryotic cytosolic ribosomes. Like bacterial ribosomes, chloroplast and mitochondrial ribosomes use a formylated tRNA. In addition, they are sensitive to many of the inhibitors of protein synthesis in bacteria. 

Most of the reactions involved in lipid biosynthesis appear to be located in the smooth endoplasmic reticulum (SER), although several enzyme activities have also been detected in the Golgi complex. Because each enzyme is a membrane-associated protein with its active site facing the cytoplasm, the biosynthesis of phospholipid occurs at the interface of ER membrane and cytoplasm. The fatty acid composition of phospholipids changes somewhat after their synthesis. (Typically, unsaturated fatty acids replace the original saturated fatty acids incorporated during synthesis.) Most of this remodeling is accomplished by certain phospholipases and acyl transferases. Presumably, this process allows a cell to adjust the fluidity of its membranes. 

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