PERSPECTIVE Biology and biochemistry of cholesterol | Ira Tabas, Series Editor SERIES INTRODUCTION Cholesterol in health and disease Ira Tabas Department of Medicine, and Department of Anatomy and Cell Biology, Columbia University, New York, New York, USA J. Clin. Invest. 110:583–590 (2002). doi:10.1172/JCI200216381. Imagine if an excess amount of a critical, life-sustain- by increasing the degree of saturation of the fatty acyl ing molecule like ATP were, by a perverse series of moieties of membrane phospholipids. However, unsat- events involving the lifestyle of modern humans, urated fatty acids in these phospholipids, particularly in causally related to a major human disease. The thought the sn-2 position, are needed for a wide variety of cellular of “ATP” being synonymous with poor health and poor signaling functions, and so an exogenous “stiffening fac- living in the minds of the lay public and press, even of tor” was needed. This factor would have to be able to health care providers, would be difficult for any self- pack tightly with the long saturated and unsaturated respecting scientist to accept. So one might view the fatty acyl chains of membrane phospholipids through biomedical history of cholesterol — indeed, this histo- van der Waals interactions. This would require a long, ry might be seen as even stranger than the hypothetical flat, and properly shaped hydrophobic molecule, accom- ATP scenario, given that evolution has devoted close to panied by additional features (see below) to help further 100 genes to the synthesis, transport, metabolism, and stabilize this interaction. regulation of cholesterol. This structurally fascinating To meet these needs, nature began with molecules, lipid is utterly essential to the proper functioning of probably originating from prebiotic anaerobic times, cells and organisms. Cholesterol, cholesterol metabo- that were formed by the sequential condensation of the lites, and immediate biosynthetic precursors of choles- two-carbon acetate molecule. Prokaryotes evolved terol play essential roles in cellular membrane physiol- enzymes to synthesize more complex linear molecules ogy, dietary nutrient absorption, reproductive biology, from these precursors (e.g., carotenoids), which were stress responses, salt and water balance, and calcium able to meet the membrane-organizing requirements metabolism. Indeed, many of the articles in this Per- of most of these primitive organisms. These primitive spective series are devoted to the normal physiology of molecules, however, lacked the proper shape, rigidity, cholesterol. Still, there is little doubt that the disease and amphipathic nature to properly organize the mem- process responsible for the leading cause of death in branes of more advanced organisms and, interestingly, the industrialized world — atherosclerosis — is a disor- two species of bacteria (see below) (1). At some point in der in which an excess of cholesterol is a major culprit. the evolution of the earliest eukaryotes or their nearest How did evolution come up with a molecule that is Archaeal relatives, the largest of these linear molecules critical for so many aspects of normal physiology, and was cyclized to form steroids, planar structures con- what went “wrong”? taining four rings (2). This shape change conferred rigidity and altered the molecule’s ability to pack with- The physiology of cholesterol in the fatty acyl tails of neighboring phospholipids. Cholesterol and cellular membranes: an evolutionary perspec- Additional features were added that helped stabilize tive. As organisms became more complex, cells required these lateral interactions, including the introduction of membranes that would provide the proper conforma- a C-C double bond (3) and of an –OH group, which tional environment for a wide variety of integral mem- converted the steroid to a sterol; the precise role of the brane proteins, such as channels, transporters, and latter modification is controversial (1). enzymes. Moreover, the cells of these advanced organ- These molecular transformations correspond to the isms required sophisticated signaling machinery, and initial conversion of acetate into acyclic, apolar squalene, this machinery would have to be organized as multipro- followed evolutionarily by the oxygenation and cycliza- tein complexes in focal, nonhomogenous areas of cellu- tion of squalene to lanosterol (Figure 1). Lanosterol is lar membranes. Put simply, these requirements were met then converted to cholesterol by oxidative demethyla- by focally increasing the “stiffness” or viscosity of phos- tion. As referred to above, two species of bacteria, Methy- pholipid bilayers. In theory, this goal could be achieved lococcus capsulatus and Mycoplasma species, require sterols for growth; while Mycoplasma obtains these sterols from the cells of higher organisms, M. capsulatus actually syn- Address correspondence to: Ira Tabas, Department of Medicine, thesizes sterols (1). In both cases, however, the structur- Columbia University, 630 West 168th Street, New York, New York 10032, USA. Phone: (212) 305-9430; Fax: (212) 305-4834; al requirements of these sterols are more lax than those E-mail: [email protected]. described above for eukaryotes (1). Interestingly, the Conflict of interest: No conflict of interest has been declared. cyclization of squalene and the conversion of lanosterol The Journal of Clinical Investigation | September 2002 | Volume 110 | Number 5 583 Figure 1 Biosynthesis of steroids in various species. Adapted from ref. 43 with permission of the editors and publisher. to cholesterol require molecular oxygen and could there- lanosterol is already a cyclic 3β alcohol. In fact, lanos- fore occur only after the evolution of aerobic cells. How- terol and other methylated derivatives cannot substi- ever, in a fascinating “evolutionary preview” of some of tute for cholesterol in mammalian cell mutants that these features, acidophilic bacteria evolved enzymes to are auxotrophic for sterols (6). Why? The three methyl anaerobically synthesize cyclic membrane-organizing groups that are removed from lanosterol to form cho- lipids called hopanoids from squalene (Figure 1). Haines lesterol are all on the so-called α-face of the molecule has speculated that hopanoids fill a unique role in aci- and thus protrude from this otherwise flat surface (Fig- dophile membrane structure, namely, the prevention of ure 2a). This is particularly true of the axial 14α-methyl inward leakage of protons (4). group of lanosterol. Bloch and others have suggested The structural requirements outlined above demand that the removal of these three methyl groups allows precision. For example, a molecule almost identical to proper fitting of the ring structure to the fatty acyl cholesterol but with a 3α-hydroxyl group instead of a chains of membrane phospholipids, particularly those 3β-hydroxyl group (“epicholesterol”) cannot function that are saturated (1, 7) (Figure 2b). According to this properly in biological membranes (5). Moreover, the model, cholesterol optimally binds natural membrane conversion of lanosterol to cholesterol involves a com- phospholipids, which usually contain a saturated fatty plex series of 18 enzymatic reactions, even though acid at sn-1 and an unsaturated fatty acid at sn-2. In par- 584 The Journal of Clinical Investigation | September 2002 | Volume 110 | Number 5 ticular, its flat α-face is ideally suited to interact with hon [16], this series). The formation of cholesterol-rich the sn-1 saturated fatty acyl group, whereas the methy- microdomains in the plasma membrane is also critical lated β-face is optimal for interaction with the more for the budding of clathrin-coated pits (17), a key step flexible unsaturated fatty acyl chains in the sn-2 posi- in receptor-mediated endocytosis. The influence of tion of an adjacent phospholipid molecule. The α-face cholesterol on normal cellular physiology is clearly of cholesterol, but not lanosterol, has the additional demonstrated by what happens when cells are experi- feature of a C-C double bond, which further enhances mentally depleted of cholesterol or when, by accidents interaction with neighboring phospholipids (3). In this of nature, mutations prevent the proper synthesis or context, Mouritsen and colleagues (8) have used deu- trafficking of cholesterol. The specific effects of cho- terium nuclear magnetic resonance spectroscopy on lesterol biosynthetic mutations are covered by Porter model membranes to demonstrate the advantage of (18) in this series, and the Perspectives by Maxfield and cholesterol over lanosterol in the formation of liquid- Wüstner (19) and by Jefcoate (20) will discuss mecha- ordered membrane domains, or “rafts” (see below). nisms of and mutations in specific aspects of intracel- Finally, the side chain of cholesterol is saturated and lular cholesterol trafficking. relatively unsubstituted with extra methyl groups, The critical membrane-organizing functions of cho- resulting in a hydrophobic structure that is usually lin- lesterol justify the evolution of an elaborate feedback ear and aligned with the plane of the steroid nucleus regulatory system. As described by Horton, Goldstein, (Figure 2a). This design allows optimal van der Waals and Brown in a recent Spotlight in the JCI (21), sterol interactions with phospholipid fatty acyl groups while starvation induces the translocation of SREBP-SCAP avoiding disruption of the bilayer (1, 7). Interestingly, complex to the Golgi apparatus, where two proteolytic plants evolved a different