Changes in Plasma Membrane Properties and Phosphatidylcholine Subspecies of Insect Sf9 Cells Due to Expression of Scavenger Receptor Class B, Type I, and CD36

Parathath, Saj; Connelly, Margery A.; Rieger, Robert; Klein, S.; Abumrad, Nada A.; Llera-Moya, Margarita de la; Iden, Charles R.; Rothblat, George H.; Williams, David L.

doi:10.1074/jbc.m404952200

Cited by 47 publications

(55 citation statements)

References 65 publications

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“…Despite the more 'rigid' environment of cholesterol-rich microdomains, experiments performed in vitro with purifi ed cholesterol-rich and -poor microdomains show that cholesterol transfers into and out of cholesterolrich microdomains much more rapidly than into/out of that while cholesterol increased the rigidity of the acyl chain region, it conversely increased the mobility/fl uidity of the polar head group region ( 40 ). The localization of the dansyl group of DChol in a less ordered microenvironment in cholesterol-rich than -poor microdomains may account for the fact that cholesterol in cholesterolrich microdomains exhibits faster and greater: i) spontaneous cholesterol traffi cking ( 6,22,26,52 ), ii) accessibility to cholesterol oxidase ( 8 ), iii) accessibility to the intracellular cholesterol transporter SCP-2 ( 6, 26, 36, 52 ), iv) direct interactions with caveolin-1, which is enriched in cholesterol-rich microdomains/caveolae, but not cholesterol-poor microdomains ( 60,61 ), v) accessibility to plasma membrane cholesterol-rich microdomains/caveolae proteins involved in cholesterol uptake/effl ux including caveolin-1, ABCA1 transporter, and SRB1 (62)(63)(64), and vi) accessibility to HDL and apoA1 ( 27,63 ). These data suggest that not only the presence of reverse cholesterol transport proteins in cholesterol-rich microdomains and the expression of intracellular cholesterol binding proteins but also the structural properties of cholesterol therein play important role(s) in regulating cholesterol uptake/effl ux.…”

Section: Discussionmentioning

confidence: 99%

Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells

Huang

McIntosh

Atshaves

et al. 2010

Journal of Lipid Research

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Section: Discussionmentioning

confidence: 99%

Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells

Huang

McIntosh

Atshaves

et al. 2010

Journal of Lipid Research

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“…Previously, SRB in ixodid ticks (Haemaphysalis longicornis) was found to play a key role in granulocyte-mediated phagocytosis to invading Escherichia coli and contributed to the first-line host defence against various pathogens [50]. Also, other studies on SRB in insects revealed the critical roles of this protein in cellular lipid regulation [51] and uptake of dietary carotenoids [52].…”

Section: Discussionmentioning

confidence: 99%

Protein-protein interactions between A. aegypti midgut and dengue virus 2: two-hybrid screens using the midgut cDNA library

Tham

Balasubramaniam

Chew

et al. 2015

J Infect Dev Ctries

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Introduction: Dengue virus (DENV) is principally transmitted by the Aedes aegypti mosquito. To date, mosquito population control remains the key strategy for reducing the continuing spread of DENV. The focus on the development of new vector control strategies through an understanding of the mosquito-virus relationship is essential, especially targeting the midgut, which is the first mosquito organ exposed to DENV infection. Methodology: A cDNA library derived from female adult A. aegypti mosquito midgut cells was established using the switching mechanism at the 5' end of the RNA transcript (SMART), in combination with a highly potent recombination machinery of Saccharomyces cerevisiae. Gal4-based yeast two-hybrid (Y2H) assays were performed against DENV-2 proteins (E, prM, M, and NS1). Mammalian two-hybrid (M2H) and double immunofluorescence assays (IFA) were conducted to validate the authenticity of the three selected interactions.Results: The cDNA library was of good quality based on its transformation efficiency, cell density, titer, and the percentage of insert size. A total of 36 midgut proteins interacting with DENV-2 proteins were identified, some involved in nucleic acid transcription, oxidoreductase activity, peptidase activity, and ion binding. Positive outcomes were obtained from the three selected interactions validated using M2H and double IFA assays. Conclusions: The identified proteins have different biological activities that may aid in the virus replication pathway. Therefore, the midgut cDNA library is a valuable tool for identifying DENV-2 interacting proteins. The positive outcomes of the three selected proteins validated supported the quality of the cDNA library and the robustness of the Y2H mechanisms.

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“…While there are several known HDL receptors in hepatocytes (SRB1, CD36, and glycosylphosphatidylinositol-anchored HDL-binding protein-1), SRB1 accounts for most of the selective uptake of cholesterol (9,27,39). Although apoA1 is known to be secreted by hepatocytes and facilitate the ABCA-1-mediated efflux of phospholipid and cholesterol from peripheral cells, apoA1 did not enhance NBD-cholesterol efflux from cultured primary hepatocytes, even though the hepatocyte plasma membrane expresses high amounts of ABCA-1 transporter (4).…”

Section: Discussionmentioning

confidence: 99%

Effect of sterol carrier protein-2 gene ablation on HDL-mediated cholesterol efflux from cultured primary mouse hepatocytes

Storey¹,

Atshaves²,

McIntosh³

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

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. Effect of sterol carrier protein-2 gene ablation on HDLmediated cholesterol efflux from cultured primary mouse hepatocytes. Am J Physiol Gastrointest Liver Physiol 299: G244-G254, 2010. First published April 15, 2010 doi:10.1152/ajpgi.00446.2009.-Although HDL-mediated cholesterol transport to the liver is well studied, cholesterol efflux from hepatocytes back to HDL is less well understood. Real-time imaging of efflux of 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-amino)-23,24-bisnor-5-cholen-3␤-ol (NBD-cholesterol), which is poorly esterified, and [ 3 H]cholesterol, which is extensively esterified, from cultured primary hepatocytes of wild-type and sterol carrier protein-2 (SCP-2) gene-ablated mice showed that 1) NBD-cholesterol efflux was affected by the type of lipoprotein acceptor, i.e., HDL3 over HDL2; 2) NBD-cholesterol efflux was rapid (detected in 1-2 min) and resolved into fast [half time (t1/2) ϭ 2.4 min, 6% of total] and slow (t 1/2 ϭ 26.5 min, 94% of total) pools, consistent with protein-and vesicle-mediated cholesterol transfer, respectively; 3) SCP-2 gene ablation increased efflux of NBD-cholesterol, as well as [3 H]cholesterol, albeit less so due to competition by esterification of [ 3 H]cholesterol, but not NBD-cholesterol; and 4) SCP-2 gene ablation increased initial rate (2.3-fold) and size (9.7-fold) of rapid effluxing sterol, suggesting an increased contribution of molecular cholesterol transfer. In addition, colocalization, double-immunolabeling fluorescence resonance energy transfer, and electron microscopy, as well as cross-linking coimmunoprecipitation, indicated that SCP-2 directly interacted with the HDL receptor, scavenger receptor class B type 1 (SRB1), in hepatocytes. Other membrane proteins in cholesterol efflux [SRB1 and ATP-binding cassettes (ABC) A-1, ABCG-1, ABCG-5, and ABCG-8] and several soluble/vesicle-associated proteins facilitating intracellular cholesterol trafficking (StARDs, NPCs, ORPs) were not upregulated. However, loss of SCP-2 elicited twofold upregulation of liver fatty acid-binding protein (L-FABP), a protein with lower affinity for cholesterol but higher cytosolic concentration than SCP-2. Ablation of SCP-2 and L-FABP decreased HDL-mediated NBD-cholesterol efflux. These results indicate that SCP-2 expression plays a significant role in HDL-mediated cholesterol efflux by regulating the size of rapid vs. slow cholesterol efflux pools and/or eliciting concomitant upregulation of L-FABP in cultured primary hepatocytes. confocal imaging; bile; scavenger receptor B1 EXCESS CHOLESTEROL IS PRIMARILY removed from the body via the liver into bile, and Ͼ95% of excreted cholesterol originates from lipoproteins, primarily HDL (9). HDL removes unesterified (free) cholesterol from tissues, transports free (as well as esterified) cholesterol to the liver, and binds to scavenger receptor class B type 1 (SRB1) at hepatocyte basolateral plasma membranes (9). Hepatic clearance of HDL free cholesterol from blood is rapid [half time (t 1/2 ) ϭ 3 min] (15, 31), consistent with very ra...

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Changes in Plasma Membrane Properties and Phosphatidylcholine Subspecies of Insect Sf9 Cells Due to Expression of Scavenger Receptor Class B, Type I, and CD36

Cited by 47 publications

References 65 publications

Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells

Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells

Protein-protein interactions between A. aegypti midgut and dengue virus 2: two-hybrid screens using the midgut cDNA library

Effect of sterol carrier protein-2 gene ablation on HDL-mediated cholesterol efflux from cultured primary mouse hepatocytes

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