Browsing by Subject "Cholesterol"
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Item Alzheimer's Disease and disordered cholesterol metabolism another illness to treat with statins?(2000-06-22) Dietschy, John M.Item Approach to cholesterol management: 2001 national cholesterol guidelines(2002-02-21) Grundy, Scott M.Item A Bacterial Cholesterol Sensor to Assess Cholesterol Accessibility in Red Blood Cells(2016-01-19) Chakrabarti, Rima Shah; Radhakrishnan, Arun; Cohen, Jonathan C.; Hobbs, Helen H.Mammals are able to gain cholesterol from two sources: diet and endogenous synthesis. However, the only means of cholesterol removal is reverse cholesterol transport (RCT), in which cholesterol is transported to the liver and exported into bile. While high density lipoprotein (HDL) is considered to be the major conduit for RCT, studies with HDL-deficient animals reveal no defect in tissue cholesterol balance. We hypothesize that red blood cells (RBCs), which contain 50% of blood cholesterol, also play a role in RCT. To measure accessible cholesterol in RBCs, we developed an assay that utilizes the cholesterol binding properties of the toxin Anthrolysin-O (ALO). We purified and fluorescently labeled domain 4 of ALO (fALOD4). We then incubated fALOD4 with RBCs from 164 subjects and measured fluorescence intensity using flow cytometry. Both intra-assay and intra-individual variability of the assay were less than 10%. In the test population, fALOD4 binding varied 10-fold. fALOD4 binding did not correlate with total RBC cholesterol but did correlate with RBC phosphatidylcholine (PC) (-0.42, p=6e-7) and lyso-phosphatidylcholine (LPC) (0.40, p=6e-6). Increasing the LPC:PC ratio in RBCs with phospholipase A2 (PLA2) increased fALOD4 binding by 3-fold. fALOD4 binding also correlated with plasma HDL (0.30, p=6e-4) and triglycerides (-0.57, p=2e-12). These data suggest that RBC accessible cholesterol varies in a population, is driven by intrinsic RBC phospholipid composition and interacts with known cholesterol transporters in the blood. Future studies will determine if variability in fALOD4 binding is driven by non-lipid RBC membrane components, is genetically determined, or contributes to atherosclerosis.Item Causes of high blood cholesterol: implications for treatment(1991-02-07) Grundy, Scott M.Item Cholesterol Accessibility in Membranes(2019-03-19) Endapally, Shreya; Alto, Neal; Radhakrishnan, Arun; Rizo-Rey, José; Blount, PaulCholesterol levels in mammalian cells are tightly regulated to lie within narrow limits. This regulation is achieved by employing multiple feedback mechanisms to regulate both synthesis and uptake of cholesterol. Most of a cell's cholesterol (~80 % of total) is in the plasma membrane (PM), but the protein machinery that senses and regulates cellular cholesterol resides in the endoplasmic reticulum (ER) membrane, which contains a very small fraction (~1% of total) of a cell's cholesterol. A carefully regulated lipid transport pathway between PM and ER allows cholesterol sensors in ER to monitor the cholesterol content of cholesterol-rich PM. This transport depends on the interactions of cholesterol with various phospholipids that control its accessibility for transport to the ER. Cholesterol in PM is organized into three different pools. One pool is accessible for transport to the ER, a second pool is sequestered by sphingomyelin (SM) and can be released by treatment with sphingomyelinase, and a third pool remains sequestered even after sphingomyelinase treatment. Here, I describe our work in developing and characterizing tools to study these different pools. The three pools were identified using bacterial toxins called cholesterol dependent cytolysins (CDCs) which selectively bind to the accessible pool of cholesterol. One example of these toxins is Anthrolysin O (ALO). To better understand the dynamics of accessible cholesterol sensing at a molecular level, we developed a stable construct of the cholesterol sensing domain of ALO (ALOD4) and performed NMR and other biophysical studies using cholesterol containing model membranes. We were able to identify residues that are significantly affected by the interaction of ALOD4 to membranes. We also developed a highly specific sensor for the SM-sequestered pool of cholesterol. This sensor is derived from a fungal toxin, Ostreolysin A (OlyA). Using X-ray crystallography, we studied the interaction of OlyA with SM and cholesterol at the atomic level. This structural analysis combined with detailed mutagenesis led us to a single point mutation in OlyA that abolishes its cholesterol specificity while retaining SM specificity. Comparing the X-ray structures of these two versions of lipid bound OlyA combined with ligand docking simulations revealed two distinct conformations of SM: one in complex with cholesterol and one free from cholesterol. Studies in live cells using OlyA and ALOD4 show that the pool of SM/cholesterol complexes in plasma membrane is maintained at a constant level across a large range of cholesterol concentrations. The development of new tools for specific forms of cholesterol (ALOD4 and OlyA) has allowed us to evaluate long-standing hypotheses regarding lipid organization in PMs and has also shed new light on lipid dynamics in the context of cellular signaling.Item Does a low plasma cholesterol concentration cause madness?(1994-03-03) Dietschy, John M.Item Evaluating the Mechanisms of 2-Hydroxypropyl-β-Cyclodextrin and Liver X Receptor Agonists as Potential Therapies for Niemann-Pick Type C Disease(2013-02-12) Taylor, Anna Marie 1986-; Mendelson, Carole R.; Kliewer, Steven A.; Goodman, Joel M.; Repa, Joyce J.Cholesterol is essential to life; therefore, the synthesis, entry, and efflux of cholesterol are tightly regulated. In some rare disease states, such as in Niemann-Pick Type C, cholesterol balance is lost leading to detrimental effects. In Niemann-Pick Type C, mutations in either of the cholesterol trafficking proteins, NPC1 or NPC2, lead to the entrapment of unesterified cholesterol within the lysosome. The accumulated cholesterol promotes increased inflammation and apoptosis throughout the body resulting in premature death, which typically occurs during adolescence in humans. Currently there are no therapies proven to halt the progression of Niemann-Pick Type C in patients; however, two separate compounds (an LXR agonist and a cyclodextrin) have been shown to increase lifespan in the Npc1-/- mouse model. Although both of these potential therapies are known to alter cholesterol dynamics in the cell, the molecular mechanism(s) through which they are able to correct the defect in Niemann-Pick Type C and ultimately to enhance survival have not been fully elucidated, which is the goal of this work. As Abcg1 is a LXR target gene and a potential mechanism through which cholesterol is trafficked from Npc1-/- cells after LXR agonist treatment, the Npc1-/-Abcg1-/- mouse was generated and evaluated. These mice die significantly earlier than Npc1-/- littermates and suggest that ABCG1 plays a vital role in reducing inflammation in Niemann-Pick Type C. In addition, comprehensive studies were done within 24 hours of cyclodextrin administration in Npc1-/- mice. These results show that cyclodextrin works in Npc1-/- mice by freeing the trapped cholesterol from the lysosome of each cell very rapidly and then releasing the cholesterol intracellularly for normal sterol processing. Finally, Npc1-/- mice were treated in combination with the LXR agonist and cyclodextrin to test if dual treatment had an additive effect on relieving Niemann-Pick Type C disease progression. The combination therapy had no further benefit over cyclodextrin alone, which implies that the two agents are acting by similar mechanism(s). Overall, this work further clarifies the molecular mechanism(s) of LXR agonists and cyclodextrins in Niemann-Pick Type C disease progression, which could lead to the development of more effective therapies for patients.Item HDL Phospholipid and ABCA1-Mediated Cholesterol Efflux Are Reduced in Patients with Very High HDL-C Who Develop Early Coronary Artery Disease(2014-04-15) Agarwala, Anandita; Khera, Amit; Hobbs, Helen H.; Grundy, Scott M.BACKGROUND: Plasma levels of high-density lipoprotein cholesterol (HDL-C) are strongly inversely associated with coronary artery disease (CAD), and high HDL-C is generally associated with apparent ‘protection’ from CAD. OBJECTIVE: We identified a number of individuals with high HDL-C levels who develop CAD, a paradoxical phenotype and hypothesized that such individuals may have HDL with altered structure and function, and compared controls with similarly high HDL-C and no coronary disease. METHODS: 55 subjects with HDL-C above the 90th percentile, early CAD, and no major known risk factors for coronary disease were identified. We selected 120 controls without CAD, each matched for race, gender, and HDL-C level. RESULTS: Comparison of HDL particle characteristics between cases and controls demonstrated a significant reduction in HDL phospholipid composition and cholesterol efflux capacity in cases as compared to controls. CONCLUSION: Reduced cholesterol efflux capacity in cases with elevated HDL-C and CAD may explain the development of early coronary artery disease. Cholesterol efflux capacity may in fact be a better predictor of the risk of coronary disease then HDL-C levels alone. The reduction in HDL phospholipid in the cases may help account for impaired cholesterol efflux.Item The Hydrophobic Handoff Between NPC2 and the N-Terminal Domain of NPC1 in the Export of Cholesterol from Lysosomes(2013-05-31) Wang, Michael Leechun; Brown, Michael S.; Goldstein, Joseph L.; Thomas, Philip J.; Roth, Michael G.; Hofmann, Sandra L.; Liang, GuoshengLow density lipoproteins (LDL) and related plasma lipoproteins deliver cholesterol to cells by receptor-mediated endocytosis. The lipoprotein is degraded in late endosomes and lysosomes where its cholesterol is released. Egress of cholesterol from late endosomes and lysosomes (hereafter referred to as lysosomes) requires two proteins: Niemann-Pick C2 (NPC2), a soluble protein of 132 amino acids; and NPC1, an intrinsic membrane protein of 1278 amino acids and 13 postulated membrane-spanning helices that span the lysosomal membrane. Recessive loss-of-function mutations in either NPC2 or NPC1 produce NPC disease, which causes death in childhood owing to cholesterol accumulation in lysosomes of liver, brain, and lung. Consistent with their cholesterol export role, NPC2 and NPC1 both bind cholesterol. The cholesterol binding site on NPC1 is located in the NH2-terminal domain (NTD), which projects into the lysosomal lumen. This domain, designated NPC1(NTD), can be expressed in vitro as a soluble protein of 240 amino acids that retains cholesterol binding activity. This thesis studies NPC2 and NPC1(NTD) in detail as summarized below. Two major differences exist between the cholesterol binding of NPC2 and NPC1(NTD). 1) Competitive binding studies and crystal structures indicate that the two proteins bind cholesterol in opposite orientations. NPC2 binds the iso-octyl side chain, leaving the 3ß hydroxyl exposed, whereas NPC1 binds the 3ß-hydroxyl, leaving the side chain partially exposed. 2) Kinetic studies of cholesterol binding reveal that NPC2 binds and releases cholesterol rapidly (half-time < 2 min at 4oC), while NPC1(NTD) binds cholesterol very slowly (half-time > 2 hr at 4oC). Its rapid cholesterol binding allows NPC2 to transfer cholesterol to and from liposomes. Unlike NPC2, NPC1(NTD) cannot rapidly transfer its bound cholesterol to liposomes. However, NPC1(NTD) can accomplish this delivery when NPC2 is present. Furthermore, cholesterol binding to NPC1(NTD) is accelerated by >15-fold when the sterol is first bound to NPC2 and then transferred to NPC1(NTD). These data led us to advance a model in which NPC2 can mediate bi-directional transfer of cholesterol to or from NPC1(NTD). In cells, we envision that NPC2 accepts cholesterol in the lysosomal lumen and transports it to membrane-bound NPC1, thus accounting for the requirement for both proteins for lysosomal cholesterol export. Amino acid residues important or binding or transfer of cholesterol on NPC2 and NPC1(NTD) were identified through alanine scan mutagenesis. For both NPC2 and NPC1(NTD), residues that decreased binding mapped to areas surrounding the binding pockets on the crystal structures; residues that decreased transfer, but not binding, mapped to discrete surface patches near the opening of the binding pockets. These surface patches may be sites where the two proteins interact to transfer cholesterol. The most severe mutations disrupting binding were P120S for NPC2 and P202A/F203A for NPC1(NTD); and those that disrupted transfer were V81D for NPC2 and L175Q/L176Q for NPC1(NTD). Furthermore, the functional significance of both the binding and transfer of cholesterol by NPC2 and NPC1(NTD) in the egress of cholesterol from lysosomes was confirmed. The above binding- or transfer-defective mutants of NPC2 and NPC1 were unable to rescue LDL-stimulated cholesteryl ester synthesis in NPC2 or NPC1-deficient cells, respectively, in contrast to wild-type NPC2 and NPC1. With these data, we envision that NPC2 binds cholesterol the instant that it is released from LDL, either as the free sterol or after cleavage of lipoprotein-derived cholesteryl esters by lysosomal acid lipase. This binding would prevent cholesterol from crystallizing in the lysosomal lumen. According to the model, NPC2 can transfer its bound cholesterol to NPC1(NTD) directly, thus avoiding the necessity for the insoluble cholesterol to transit the water phase. This transfer of cholesterol from NPC2 to NPC1(NTD) has a special functional relevance in light of the near-absolute insolubility of cholesterol in water, and we have named this process a "hydrophobic handoff."Item Impaired Cholesterol Efflux Capacity May Help Explain Development of Early Coronary Artery Disease in Subjects with Very High HDL-C(2014-02-04) Agarwala, Anandita; Rodrigues, Amrith; Trinidade, Kevin; Risman, Marjorie; Qu, Liming; Cuchel, Marina; Billheimer, Jeffrey; Rader, Daniel J.Plasma levels of high-density lipoprotein cholesterol (HDL-C) are strongly inversely associated with coronary artery disease (CAD), and high HDL-C is generally associated with apparent 'protection' from CAD. A minority of individuals with very high HDL-C levels also develops CAD, a paradoxical phenotype. We hypothesize that such individuals may have HDL with altered structure and/ or function, and compared these individuals (cases) to individuals with very high HDL-C without CAD (controls). We identified 55 subjects with HDL-C above the 90th percentile, early CAD, and no major risk factors for coronary disease. We selected 120 controls without CAD, each matched for race, gender, and HDL-C level. Controls were selected to be the same age or no more than 10 years older than the cases. Studies to assess HDL composition and size distribution, cholesterol efflux capacity, and lecithin-cholesterol acyltransferase (LCAT) activity in cases and controls were conducted. Comparison of HDL particle characteristics between cases and controls demonstrated a significant reduction in HDL phospholipid composition between cases and controls (92 ± 37 mg/dl vs. 109 ± 43 mg/dL, p value 0.0095). The mean plasma total cholesterol efflux capacity was significantly reduced in subjects with elevated HDL-C and CAD as compared to controls (1.96 ± 0.39 % efflux/ 2hr/ 1% plasma vs. 2.11 ± 0.43 % efflux/ 2hr/ 1% plasma, p value 0.040). The reduction became even more significant when looking at mean ABCA1- selective cholesterol efflux between cases and controls 0.60 ± 0.24 % efflux/ 2hr/ 1% plasma vs. 0.71 ± 0.32 % efflux/ 2hr/ 1% plasma, p value 0.033). Furthermore, there was a significant reduction in mean efflux per HDL particle in cases as compared to controls (0.023 ± 0.005 % efflux/ 2hr/ 1% plasma vs. 0.025 ± 0.006 % efflux/ 2hr/ 1% plasma, p value 0.029). No significant difference was observed between cases and controls in HDL particle size or plasma LCAT activity. Reduced cholesterol efflux capacity in cases with elevated HDL-C and CAD may explain the development of early coronary artery disease. This finding reinforces the belief that cholesterol efflux capacity may in fact be a better predictor of the risk of coronary disease then HDL-C levels alone. Furthermore, the reduction in HDL phospholipid in the cases may help account for impaired cholesterol efflux.Item Insig-Mediated Regulation of Mammalian HMG COA Reductase Ubiquitnation and Degradation(2004-12-15) Sever, Navdar; Brown, Michael S.3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase (HMGR) catalyzes the conversion of HMG CoA to mevalonate, which is the rate limiting step in the production of cholesterol and numerous nonsterol isoprenoid products. Mammalian HMGR is regulated by transcriptional and post-transcriptional feedback mechanisms. The transcriptional regulation is mediated by sterol regulatory element binding proteins (SREBPs), which are synthesized as inactive precursors in the endoplasmic reticulum (ER) membrane. In the absence of sterols, SREBP cleavage activating protein (SCAP) escorts SREBPs from ER to the Golgi apparatus, where SREBPs are cleaved by site 1 and site 2 proteases so as to release their amino terminal transcription factor domains to the nucleus. Sterols inhibit the exit of SCAP-SREBP complex from the ER by promoting the binding of two related polytopic ER membrane proteins, Insig-1 and Insig-2, to the membrane domain of SCAP. Insig-1, but not Insig-2, is an SREBP target gene, causing Insig-1 levels to drop in the presence of sterols, when it is expected to exert its action. The degradation of HMGR requires both sterols and a nonsterol product of the mevalonate pathway and the eight membrane spanning segments in its amino terminus. The membrane domains of HMGR and SCAP bear sequence similarity prompting the investigation of whether Insig proteins can also bind to HMGR. Indeed, Insig-1 and Insig-2 were found to interact with HMGR in a regulated manner and mediate its proteasomal degradation. This effect can be specifically inhibited by overexpressing the membrane domain of SCAP. Insigs were shown to promote the ubiquitination of HMGR on lysine 248 in the cytoplasmic loop between transmembrane segments 6 and 7. In an attempt to achieve a better understanding of the mechanism by which HMGR is degraded, a genetic approach was developed to select mutant somatic cells that cannot degrade HMGR in the presence of sterols. The isolation and characterization of Chinese hamster ovary cells deficient in Insig-1 confirmed the endogenous requirement of Insig-1 for HMGR degradation and revealed the role of differential regulation of Insig-1 and Insig-2 in terms of SREBP processing. These studies revealed a complex feedback regulatory system governing cholesterol homeostasis.Item Lipid management strategies for increasingly ambitious goals(2006-09-29) Abate, NicolaItem Lowering plasma cholesterol by raising LDL receptors(1981-08-06) Brown, Michael S.Item Mechanistic Dissection of Insig-1, a Master Regulator of Cholesterol Homeostasis(2006-05-15) Gong, Yi; Brown, Michael S.Insigs are polytopic membrane proteins of the endoplasmic reticulum (ER) that regulate lipid synthesis by controlling the sterol-mediated vesicular transportation of sterol regulatory element binding proteins (SREBPs). SREBPs are ER bound transcription factors that form complexes with Scap. In sterol-depleted cells, Scap escorts SREBPs from the ER to the Golgi apparatus, where SREBPs are proteolytically cleaved to liberate the nuclear fragments that activate genes for cholesterol synthesis and uptake. When sterols overaccumulate in cells, the Scap/SREBP complex is retained in the ER by the anchor proteins called Insigs. In this thesis I describe the formation of a complex between Insig-1 and Scap in a sterol regulated fashion which facilitates the ER retention of Scap. To understand the molecular basis of the interactions between Insig-1 and Scap, I use a site-directed mutagenesis approach to select residues in Insig-1 that are essential for Insig-1/Scap complex formation. This study reveals a functional role for the amino acid Asp-205, which is located at the beginning of the fourth loop of Insig-1. Mutation of this aspartic acid to alanine produces an inactive Insig-1 that no longer binds to Scap, and leads to sterol-resistant processing of SREBPs. Mammalian cells express two Insig proteins differ in their mode of control. Insig-1, but not Insig-2, is an SREBP target gene. Also, Insig-1 protein is degraded more rapidly than Insig-2. Thus, Insig-1 is the focus of the study. I further demonstrate that degradation of Insig-1 is regulated by sterols. When ER cholesterol content is low, Insig-1 is ubiquitinated on lysines 156 and 158 and degraded in proteasomes. Sterol-induced binding of Insig-1 to Scap prevents Insig-1 ubiquitination and degradation. The dynamic change in Insig-1 protein stability, together with its transcriptional control by nuclear SREBPs, creates a new model for the convergent inhibition of SREBP processing and cholesterol supply in animal cells. Taken together, these studies established Insig-1 as the master regulator in the cholesterol homeostasis.Item Medical therapy of cholesterol gallstone disease(1984-08-09) Dietschy, John M.Item Modalities of Cholesterol Binding and Modulation of the NPC Proteins and Scap(2011-12-14) Motamed, Massoud; Brown, Michael S.Low density lipoproteins (LDL) and related plasma lipoproteins deliver cholesterol to cells by receptor-mediated endocytosis. The lipoprotein is degraded in late endosomes and lysosomes, allowing cholesterol to be released. Export of cholesterol from late endosomes and lysosomes (hereafter referred to as lysosomes) requires two lysosomal proteins: Niemann-Pick C2 (NPC2), a soluble protein of 132 amino acids; and NPC1, a membrane protein with 13 putative membrane-spanning helices. Recessive loss-of-function mutations in either NPC2 or NPC1 produce NPC disease, which causes death owing to lipid accumulation in lysosomes of liver, brain, and lung. Consistent with their cholesterol export role, NPC2 and NPC1 both bind to cholesterol. The cholesterol binding site on NPC1 is located in the NH2-terminal domain (NTD), which projects into the lysosomal lumen. This domain, designated NPC1 (NTD), can be expressed in vitro as a soluble protein of 240 amino acids that maintains cholesterol binding activity. This thesis studies NPC2 in detail as summarized below. Despite a shared role as cholesterol binding proteins, NPC2 and NPC1 (NTD) bind to cholesterol in opposite orientations. The crystal structures of NPC2 and NPC1 (NTD) have been solved, and NPC2 binds cholesterol with the iso-octyl chain facing the interior of the protein, whereas, NPC1(NTD) binds cholesterol with the 3ß-hydroxyl facing the interior of the protein. Another striking difference is the kinetics of this cholesterol binding. NPC2 binds and releases cholesterol rapidly (half-time < 2 min at 4oC), while NPC1 (NTD) binds cholesterol very slowly (half-time > 2 hr at 4oC). However, NPC2 can stimulate the rate of cholesterol binding to NPC1 (NTD) (>15-fold in vitro). This stimulation of cholesterol binding to NPC1 (NTD) by NPC2 is believed to occur through a direct transfer of cholesterol from NPC2 to NPC1(NTD). Amino acid residues important for binding or transfer of cholesterol on NPC2 were identified through alanine scan mutagenesis. Residues that decreased binding thermodynamics and/or kinetics mapped to areas surrounding the binding pockets on the crystal structures; residues that decreased transfer, but not binding, mapped to discrete surface patches near the exposed residues of the binding pockets. These surface patches may be sites where the two proteins interact to transfer cholesterol. The most deleterious binding mutant was P120S, a residue in the cholesterol binding pocket; the most deleterious transfer mutant was V81D, a residue on the hydrophobic patch extending outward from the cholesterol binding pocket. The above mutants of NPC2 were unable to rescue LDL-stimulated cholesteryl ester synthesis in NPC2-deficient cells, in contrast to wild-type NPC2. Once LDL-derived cholesterol leaves the lysosomes, it is transported to the endoplasmic reticulum (ER), where it serves a regulatory role in cholesterol homeostasis. In the ER, these regulatory functions include activation of acetyl-coenzyme A acetyltransferase (ACAT), allowing for esterification of cholesterol for storage, and regulation of sterol regulatory element–binding protein (SREBP) localization, a transcription factor that regulates key enzymes for cholesterol synthesis. SREBP cleavage-activating protein (Scap) is the switch that controls SREBP, and therefore cholesterol synthesis. Scap senses cholesterol abundance in the ER and acts as an escort protein. In sterol depleted cells, Scap escorts SREBP to the Golgi complex, where two proteases cleave SREBP, thereby releasing its transcriptionally active domain so that it can go to the nucleus and activate transcription of genes involved in cholesterol synthesis and uptake. When cholesterol in abundant, the sterol binds to Scap and triggers a conformational change in the protein that prevents it from escorting SREBPs to the Golgi for proteolytic cleavage. Scap is a 1276 amino acid protein that consists of two domains: an N-terminal domain with 8 transmembrane spanning regions and a C-terminal domain that projects into the cytosol and associates with SREBPs. Previous studies have localized the cholesterol-binding activity of Scap to its membrane domain. Studies described in this thesis identify the cholesterol binding pocket in Scap and identify key residues that play an important role in the protein’s responsiveness to cholesterol binding. The first loop region of Scap (hereafter referred to as Scap(Loop1)) was purified as a recombinant protein and found to have cholesterol binding activity. The specificity of this sterol binding was determined through competition studies and shown to be physiologically relevant. Additionally, this binding affinity and specificity was similar to that of the membrane domain of Scap. Subsequently, alanine scan mutagenesis was performed on Scap(Loop1). Through this approach, several mutations of Scap were identified that constitutively adopt the cholesterol-bound state. This data demonstrates that Scap(Loop1) binds to cholesterol and that the binding then helps induce the conformational change required for Scap to anchor SREBP in ER membranes.Item [News](1992-02-06) Cannella, Heidi HarrisItem [News](1988-06-23) Harrell, AnnItem [News](1985-05-17) Bosler, Tommy JoyItem [News](1983-07-01) Rutherford, Susan; Williams, Ann
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