Lipid profiling of FPLC-separated lipoprotein fractions by electrospray ionization tandem mass spectrometry

Wiesner, Philipp; Leidl, Katharina; Boettcher, Alfred; Schmitz, Gerd; Liebisch, Gerhard

doi:10.1194/jlr.d800028-jlr200

Cited by 312 publications

(337 citation statements)

References 54 publications

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“…4B and Table 4). However, the phosphatidylethanolamine/phosphatidylcholine and sphingomyelin/phosphatidylcholine ratios that distinguish the lipoprotein classes 31 differed between LVPs and all other TRLs. Phosphatidylcholine was the most abundant phospholipid class in all four patients' lipoproteins and LVP.…”

Section: Lvps and Vldl Have Similar Triacylglycerolmentioning

confidence: 87%

High plasma level of nucleocapsid-free envelope glycoprotein-positive lipoproteins in hepatitis C patients

et al. 2012

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Hepatitis C virus (HCV) particles associate viral and lipoprotein moieties to form hybrid lipoviral particles (LVPs). Cell culture-produced HCV (HCVcc) and ex vivo-characterized LVPs primarily differ by their apolipoprotein (apo) B content, which is low for HCVcc, but high for LVPs. Recombinant nucleocapsid-free subviral LVPs are assembled and secreted by apoB-producing cell lines. To determine whether such subviral particles circulate in HCV-infected individuals, LVPs complexed with immunoglobulin were precipitated with protein A from low-density plasma fractions of 36 hepatitis C patients, and their lipid content, apolipoprotein profile, and viral composition were determined. HCV RNA in LVPs was quantified and molar ratios of apoB and HCV genome copy number were calculated. LVPs lipidome from four patients was determined via electrospray ionization/tandem mass spectrometry. Protein A-purified LVPs contained at least the envelope glycoprotein E2 and E2-specific antibodies. LVPs were present in every patient and were characterized by high lipid content, presence of apolipoproteins characteristic of triglyceride-rich lipoproteins (TRLs), HCV RNA, and viral glycoprotein. Importantly, save for four patients, LVPs fractions contained large amounts of apoB, with on average more than 1 3 10 6 apoB molecules per HCV RNA genome. Because there is one apoB molecule per TRL, this ratio suggested that most LVPs are nucleocapsid-free, envelope glycoprotein-containing subviral particles. LVPs and TRLs had similar composition of triacylglycerol and phospholipid classes. Conclusion: LVPs are a mixed population of particles, comprising predominantly subviral particles that represent a distinct class of modified lipoproteins within the TRL family. (HEPATOLOGY 2012;56:39-48) H epatitis C virus (HCV) is a member of the Flaviviridae family and a major cause of chronic hepatitis often leading to liver cirrhosis and hepatocellular carcinoma. 1 Chronic hepatitis C is a complex disease associated with host metabolic modifications resulting in a unique metabolic syndrome including insulin resistance, liver steatosis, and hypobetalipoproteinemia. 2,3 The most striking link between HCV and lipids resides in the association of HCV particles with lipoproteins. 4,5 Indeed, HCV virions with a density <1.06 g/mL are associated with lipoproteins, thus forming hybrid particles known as lipoviral particles (LVPs). These low-density viral particles are globular, rich in triacylglycerol and total cholesterol (TChol) and contain the viral envelope glycoproteins and nucleocapsid (composed of HCV RNA and core protein). In addition, LVPs contain all the apolipoproteins (apo) that define the triacylglycerolrich lipoproteins (TRLs). Indeed, apolipoprotein (apo) B, apoE, apoCI, apoCII, and apoCIII, all of which characterize very low-density, intermediate-density, and low-density lipoproteins (VLDL, IDL, and LDL,

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Section: Lvps and Vldl Have Similar Triacylglycerolmentioning

confidence: 87%

High plasma level of nucleocapsid-free envelope glycoprotein-positive lipoproteins in hepatitis C patients

et al. 2012

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“…LPA 1 /EDG2 was the first to be discovered in 1996, followed soon after by LPA 2 /EDG4 and LPA 3 /EDG7 [81][82][83][84] . These receptors have a ubiquitous distribution, and the other five GPCRs in the Edg-subfamily are receptors for sphingosine-1-phosphate (S1P) (S1P [1][2][3][4][5] ), which is the sphingolipid analog of LPA [80] . Knockout (KO) mice have been generated for all the known LPA receptors, as well as double KOs for combinations of LPA 1-3 and the triple KO of LPA [1][2][3] , all of which are viable [6,[85][86] .…”

Section: Lpa Receptor Diversity and Signalingmentioning

confidence: 99%

“…These receptors have a ubiquitous distribution, and the other five GPCRs in the Edg-subfamily are receptors for sphingosine-1-phosphate (S1P) (S1P [1][2][3][4][5] ), which is the sphingolipid analog of LPA [80] . Knockout (KO) mice have been generated for all the known LPA receptors, as well as double KOs for combinations of LPA 1-3 and the triple KO of LPA [1][2][3] , all of which are viable [6,[85][86] . LPA 1 deletion shows craniofacial deformity while LPA 3 deletion leads to delayed implantation of embryos and impaired embryo spacing [87][88] .…”

Section: Lpa Receptor Diversity and Signalingmentioning

confidence: 99%

“…LPA 6 /p2y5 appears to be important for hair growth since genetic truncations of the receptor leads to hypotrichosis simplex, a set of diseases involving familial hair loss [97] . In vitro, this receptor only responds to μM concentrations of LPA while LPA [1][2][3][4][5] will activate in response to nM concentrations [98] . Finally, there are at least three more proposed LPA receptors (GPR87, P2Y10 and GPR35), but their specificity for LPA requires further experimental validation [80] .…”

Section: Lpa Receptor Diversity and Signalingmentioning

confidence: 99%

“…1). Normally, LPA concentrations in plasma are about 100 nmol/L and this LPA is bound to albumin [1][2] . LPA can rise to about 1 μmol/L in serum as a result of platelet activation [3][4][5] .…”

Section: Introductionmentioning

confidence: 99%

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Recent advances in targeting the autotaxin-lysophosphatidate-lipid phosphate phosphatase axis in vivo

2016

J Biomed Res

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Extracellular lysophosphatidate (LPA) is a potent bioactive lipid that signals through six G-protein-coupled receptors. This signaling is required for embryogenesis, tissue repair and remodeling processes. LPA is produced from circulating lysophosphatidylcholine by autotaxin (ATX), and is degraded outside cells by a family of three enzymes called the lipid phosphate phosphatases (LPPs). In many pathological conditions, particularly in cancers, LPA concentrations are increased due to high ATX expression and low LPP activity. In cancers, LPA signaling drives tumor growth, angiogenesis, metastasis, resistance to chemotherapy and decreased efficacy of radiotherapy. Hence, targeting the ATX-LPA-LPP axis is an attractive strategy for introducing novel adjuvant therapeutic options. In this review, we will summarize current progress in targeting the ATX-LPA-LPP axis with inhibitors of autotaxin activity, LPA receptor antagonists, LPA monoclonal antibodies, and increasing low LPP expression. Some of these agents are already in clinical trials and have applications beyond cancer, including chronic inflammatory diseases.

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Chromatography of Membrane Proteins and Lipoproteins

Zolla

D’Alessandro

Lana

2016

Encyclopedia of Analytical Chemistry

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The available methods for the separation of membrane proteins and lipoproteins are sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE), followed by immunoblotting, isoelectric focusing (IEF), and capillary electrophoresis (CE), along with the recently introduced gel‐based native techniques (blue native (BN) and clear native (CN)), and high‐performance liquid chromatography (HPLC). In this article, it is shown that HPLC techniques, given their wide versatility, relative ease of use, and high resolution, may be considered the most valuable tool for the characterization of virtually any hydrophobic protein. Application examples are described, and comparisons with other methods are discussed. Moreover, HPLC is not a destructive technique, and therefore, proteins, once separated, are available for further analytical investigations. Among these techniques, quantitative and qualitative analyses of the separated fractions can be obtained through other biophysical approaches, such as crystallography or structural spectroscopy. Most of these approaches require preliminary protein purification (90% or higher), which could be rapidly obtained through preliminary HPLC.

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Lipid profiling of FPLC-separated lipoprotein fractions by electrospray ionization tandem mass spectrometry

Cited by 312 publications

References 54 publications

High plasma level of nucleocapsid-free envelope glycoprotein-positive lipoproteins in hepatitis C patients

High plasma level of nucleocapsid-free envelope glycoprotein-positive lipoproteins in hepatitis C patients

Recent advances in targeting the autotaxin-lysophosphatidate-lipid phosphate phosphatase axis in vivo

Chromatography of Membrane Proteins and Lipoproteins

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