S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

Discuss the physiological control of cholesterol metabolism and its dysfunction

Introduction
The understanding of cholesterol metabolism is a relatively new topic in physiological research. Some very relevant publications have been highly considered by the scientific community in the past decades; for instance, the Nobel Prize in Physiology and Medicine in 1985 for Brown and Goldsteins studies in regulation of cholesterol metabolism. Currently, scientists mainly focus on the understanding of how our body is able to control cholesterol to achieve homeostasis. Moreover, by studying the diseases where cholesterol control mechanisms are malfunctioning, we can comprehend the genetics and physiological concepts involved. Over the course of this essay, we are going to come across some basic concepts about cholesterol metabolism and functionality, its regulation and the diseases that can be developed in case of dysfunction.

Basic concepts on cholesterol metabolism

Cholesterol is a steroid molecule normally synthesised in mammalian cells of the liver or the intestine. It is transported in the blood plasma across the whole organism ready to reach any other tissue. It is an important structural component of the cell membrane and essential for permeability and fluidity. It is ideally located among saturated membrane lipids in the cell membrane. Its presence increases the ordering of these lipids restricting diffusion (therefore reducing permeability). It is also important for the production of bile acids, vitamin D and several hormones [1]. Its presence is essential for a correct cell metabolism; however failure of regulation or transportation of cholesterol can lead to cardiovascular disorders, which will be argued later on [2].

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

Cholesterol synthesis starts with the hydrolysis of one molecule of Acetyl CoA and one molecule of Acetoacetyl CoA to form HMG-CoA in the Endoplasmic Reticulum. The enzyme HMG-CoA reductase drives this reaction and an energy input is required. Immediately afterwards, several chemical reactions take place to form a molecule of cholesterol [4]. The full reaction is illustrated in Figure 1 below. 1 Acetyl CoA + 1 Acetoacetyl CoA HMG-CoA* (hydrolisis) 1 HMG-CoA + HMG-CoA reductase 1 mevalonate (reduction) 1 mevalonate + ATP! 3-isopentenyl pyrophosphate (decarboxylation) (3x) 3-isopentenyl pyrophosphate + geranyl transferase farnesyl pyrophospate (condense of 3 molecules) (2x) farnesyl pyrophosphate + squalene synthase squalene (condense of 2 molecules) 1 squalene + oxidosqualene cyclase lanosterol (cyclation) 1 lanosterol CHOLESTEROL (conversion) * HMG-CoA= also known as 3-Hydroxy-3-methylglutaryl CoA. Figure 1. Diagram of the biosynthesis of cholesterol

Moreover, we need to consider that we are talking about a negative feedback system; which upon the consumption of cholesterol, inhibits the biosynthesis of cholesterol in the body [3]. Any source of animal fat contains cholesterol, such as cheese, eggs or pork. It has been found the more cholesterol ingested in the diet, the less cholesterol is biosynthesised in cells [5]. Once cholesterol has been synthesised, it has to be available in any other spots around the organism. This is achieved by the transport of cholesterol molecules in lipoproteins across the plasma on the blood vessels, due to the hydrophobic properties of cholesterol. There are two types of lipoproteins that can transport cholesterol (with different densities of cholesterol); Low Density Lipoproteins (LDL) and High Density Lipoproteins (HDL). Cholesterol is introduced into the target cell by endocytosis. Cholesterol is then excreted by

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

the liver in the form of sterols, mostly reabsorbed and thus, recycled in the organism [6].

Physiological regulation of cholesterol in the organism

As many other compounds in the organism, cholesterol levels need to be controlled in order to avoid deficiency or excess. These can lead to health disorders. This regulation is achieved in several ways. Cholesterol homeostasis control is mainly achieved by the repression of transcription of genes that govern the biosynthesis and receptor-mediated uptake of cholesterol molecules. The SREBP protein family regulate transcription of HMG-CoA reductase and other enzymes involved in cholesterol biosynthesis; as well as regulate the LDL receptor transcription that helps supplying cholesterol to cells. They were also found to be involved in the synthesis of fatty acids and its uptake in cells. Brown and Goldstein identified three different types of SREBPs (1997). Their structure is explained in Figure 2 [7].

Figure 2. Proteomics of SREBP-1a and SREBP-2. *Note that the sequence of SREBP-1c is identical to 1a except for a domain [7].

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

For all of them, the NH2-terminal domain (green part in the diagram) is thought to be a transcription factor. When binding a certain sequence of DNA, transcription process begins. The NH2-terminal domain must be released from the membrane and enter the nucleus by a proteolytic cascade regulated by sterols, which is illustrated in Figure 3.

Cleavage at site 1

Cleavage at site 2

Protease breaks the covalent bond between the two transmembrane domains of SREBP Protease (from first transmembrane domain) clips the NH2terminal fragment releasing it into the cytosol and then entering the nucleus

At the Endoplasmic Reticulum (ER)

At the Golgi Apparatus

Figure 3. Diagram of the proteolytic cascade of SREBP. *Note: It leads to the entry of NH2 terminal of SREBP into the nucleus, thus influencing transcription.

Cleavage at site 1 is strongly dependent on the presence of sterols, however cleavage at site 2 has not been found directly regulated by these but requires previous cleavage at site 1. Biochemical structure of proteases involved in this process has not been described but they are thought to be unusual enzymes. Cellular locations are also unknown but a hypothetical scenario has been proposed [7]. SREBP activity is regulated by protein SCAP. It is remarkable that the NH2terminal domain is particularly similar to the one in HMG-CoA reductase. Thus, this domain has a sterol-sensing function: if this domain is deleted, SREBP is no longer degraded due to sterol regulation. Brown and Goldstein interpreted these data as that SCAP is a required activator of SREBP

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

cleavage and can be abolished by sterols. SCAP might bind SREBPs in respectively WD region and COOH-terminal of both macromolecules. This binding has been found to be required for site 1 proteolysis; sterols could disrupt this by blocking the binding [7]. They also described the significance of the SREBP regulation of cholesterol biosynthesis. When cholesterol levels in hamsters livers were decreased, the amount of SREBPs increased and the efficiency of the proteolytic cascade surged notably [7]. Moreover, transport of cholesterol across the cells is regulated by the activity of the LDL receptor (a cell surface protein) that binds LDL, regulating the rate at which cholesterol is transferred to the cells. LDL receptor activity is inversely proportional to the content of cholesterol in the cell. Suppression of synthesis of LDL receptors is activated by SREBP activity when there is sufficient LDL in the cell, blocking further transport into the cell and avoiding accumulation of sterols in them. Conversely, cells obtain cholesterol by increasing the quantity of LDL receptor proteins when they need to increase its presence [6].

Dysfunction of control mechanisms of cholesterol homeostasis

As we have stated before, cholesterol is essential for cell survival, nevertheless excessive quantities can cause diseases that are today considered to be some of the major causes of death in the western world. These are principally familiar hypercholesterolemia and atherosclerosis. Sometimes, metabolic processes are shown to fail due to a failure of some stage of the pathway. For instance, in some occasions cells have been found to be sterol-resistant and fail to suppress the synthesis of LDL receptors or other cells that have defective phenotypes in regulation of SREBP cleavage. These genetic disorders (mostly single gene disorders) [6] can produce dysfunction of the metabolic pathways of controlling cholesterol homeostasis

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

giving rise to decontrolled levels of cholesterol in the organism; possibly causing major clinical disorders [7]. Patients of homozygous familial hypercholesterolemia have cells that lack of LDL receptors and hence cannot take up the LDL molecules. As a consequence, LDL is not able to suppress cholesterol synthesis by stopping HMG-CoA reductase gene transcription [6]. Effects of this metabolic pathway in diseased cells and healthy ones are shown in figure 4.

Figure 4. Comparison of LDL-receptor activity in normal and homozygous familial hypercholesterolemia [6].

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

*NB: Dot = normal; Triangle= familial hypercholesterolemia.

Moreover, atherosclerosis (or hardening of the arteries) is a cardiovascular disease caused by the thickening of the arterys wall as a consequence of a fatty acids accumulation, primarily cholesterol. This obstruction in the form of plaques causes an inflammatory response promoted by LDLs. Usually these thrombi travel across the blood stream eventually occluding branches of the arteries causing thromboembolism; and sometimes can even cause a heart stroke [8].

Conclusion
We have discussed the basis of cholesterol control in mammalian organisms. The significance of genetics and enzymes has been argued, as well as the diseases caused by the failure of regulating pathways. Over the past few decades, research in cholesterol metabolism and its regulation has given scientists enough understanding in cellular biology to achieve a clinical application of their research. We have learned that regulation of LDL-receptors is essential for the right homeostasis of cholesterol. In diseased individuals this regulation is incorrect; however it can be fixed through drugs and diet, reducing the frequency of these cardiovascular diseases common in our society. There is hope that this knowledge will lead to the discovery of truly effective treatments of these pathologies in the near future.

S1036934 Cristina Moreno

Physiology 2 essay-2011 Group 7

References
1. Ikonen, E. (2008). Cellular cholesterol trafficking and compartmentalization. Nature Rev. Mol. Cell Biol. 9: 125-138. 2. Goedeke, L., Fernandez-Hernando, C. (1945). Regulation of cholesterol homeostasis. Cel. Mol. Life Sci. 1 3. Brown, M.S.; Goldstein, J.L. (1984). How LDL receptors Influence cholesterol and atherosclerosis. Scientific American. 251: 52-60. 4. Rhodes, C.l.; Stryer, L.; Tasker, R. (1995). Biochemistry (4th ed.). San Francisco: W.H. Freeman. 280-703 5. Schoenheimer, R.; F. Breusch. (1933). Synthesis and destruction of cholesterol in the organism. J. Biol. Chem. 103: 439448. 6. Brown M.S.; Goldstein J.L. (1976). Receptor-Mediated control of cholesterol metabolism. Science 16 191 (4223): 150-154. 7. Brown M.S.; Goldstein J.L. (1997) The SREBP Pathway: regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor. Cell. Journal. 89: 331-340. 8. Ross R. (1993). The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 362 (6423): 8019.