scholarly journals The isolation and characterization of high-density-lipoprotein subfractions containing apolipoprotein E from human plasma

1992 ◽  
Vol 284 (2) ◽  
pp. 477-481 ◽  
Author(s):  
H M Wilson ◽  
B A Griffin ◽  
C Watt ◽  
E R Skinner

1. Plasma high-density lipoprotein (HDL) was separated by heparin-Sepharose affinity chromatography into a non-bound, apolipoprotein E-poor, and a bound, apolipoprotein E-rich, fraction through the binding effect of Mn2+ in the column buffer. 2. The application of a series of elution buffers in which the concentration of Mn2+ was progressively replaced by Mg2+ resulted in the separation of the bound HDL into five subfractions. 3. Each subfraction migrated a different distance on gradient-gel electrophoresis. Three of the subfractions had RF (relative migration compared with BSA) values within the range of HDL2b. One subfraction contained largely HDL2a, with some material in the regions of HDL2b and HDL3a, and one subfraction spanned the RF regions of HDL2a, HDL3a and HDL3b. 4. The number of molecules, per HDL particle, of cholesteryl ester, non-esterified cholesterol and phospholipid increased with particle size, whereas triacylglycerol passed through a maximum and the number of amino acid residues remained approximately the same. 5. Apolipoprotein (apo) A-I was the major apoprotein in all five subfractions, but the latter differed appreciably in their contents of apo A-II and apo E. 6. The major fatty acid component of each subfraction was linoleic acid, with moderate amounts of C16:0 and C18:1 fatty acids and a smaller content of C18:0, C20:4,n-6 and C22:6,n-3, with no significant difference in composition between the subfractions. 7. This paper provides the first description of a method for the isolation of three subfractions of HDL2b together with other subfractions in quantities that are sufficient for further analytical or metabolic studies.

1995 ◽  
Vol 311 (2) ◽  
pp. 611-615 ◽  
Author(s):  
Y D Fragoso ◽  
E R Skinner

Apolipoprotein (apo) E plays an important role in mediating high-density lipoprotein (HDL) cholesterol transport and uptake by the liver. Evidence for and against the existence of conventional liver receptors for HDL containing apoE have been reported, although the selective uptake of the cholesterol moiety of HDL has been demonstrated. The present study investigated the hepatic uptake of subfractions of HDL separated on the basis of their apoE content. Rabbit HDL and its apoE-rich and apoE-poor subfractions, separated by heparin-Sepharose affinity chromatography, were labelled in their apoprotein moieties with [14C]sucrose and in their cholesteryl ester moiety with 3H. No binding of either subfraction to rabbit liver membranes could be detected. With cultured HepG2 cells, however, there was a high uptake of 3H but a very low uptake of 14C from both HDL subfractions, demonstrating that selective uptake was operating. Addition of unlabelled apoE-poor HDL inhibited the uptake of both labels from the two subfractions to the same extent. These studies, which differed from previously reported investigations by employing native homologous HDL subfractions of known apolipoprotein composition, demonstrated that apoE is not directly involved in the selective uptake of HDL cholesterol by the liver. In the absence of specific binding sites on liver membranes, it is suggested that an alternative mechanism might exist for the clearance of HDL cholesterol from the plasma.


1995 ◽  
Vol 41 (1) ◽  
pp. 165-169 ◽  
Author(s):  
P W Wilson

Abstract Although a variety of methods have become available for the determination of high-density lipoprotein (HDL) subfractions in plasma, a review of published data from nine studies of coronary disease outcomes and 10 investigations of coronary artery disease severity do no suggest that measurement of HDL2-cholesterol (C) and HDL3-C offer any advantage in the prediction of coronary disease over the determination of total HDL-C alone. Apolipoprotein E is typically present in plasma as one of six isoforms, the six being encoded by three common alleles, epsilon 2, epsilon 3, and epsilon 4. The epsilon 3 allele is the most common, the epsilon 4 allele has been reported to be associated with higher cholesterol concentrations, and the epsilon 2 and epsilon 4 alleles are both associated with higher triglyceride concentrations. Clinical and arteriographic studies of coronary disease suggest that vascular disease risk is increased among persons with the epsilon 4 allele.


1986 ◽  
Vol 32 (4) ◽  
pp. 646-651 ◽  
Author(s):  
J Peynet ◽  
M Fénéant-Thibault ◽  
A Legrand ◽  
D Marot ◽  
F Rousselet ◽  
...  

Abstract An abnormal high-density lipoprotein (HDL) subfraction, detected during periods of mild jaundice in the serum of seven children with chronic cholestasis from birth, was isolated and characterized. This fraction, identified by its slow alpha electrophoretic migration, is present in addition to normal HDL and differs from the abnormal HDL previously described in cholestatic syndromes. It is devoid of apolipoprotein B but is precipitated by phosphotungstate-MgCl2. These properties allowed its isolation by double selective precipitation. This subfraction is undetectable with this procedure in the serum of healthy subjects, is rich in cholesterol, and contains a large amount of apolipoprotein E, which may explain its precipitation by phosphotungstate-MgCl2. These apo E-containing HDL may play a major role in the lipid metabolism of patients with long-standing cholestasis during periods of mild jaundice.


Circulation ◽  
2018 ◽  
Vol 137 (suppl_1) ◽  
Author(s):  
Rafeal L Baker ◽  
Jean N Utumatwishima ◽  
Stephanie T Chung ◽  
Anne E Sumner

Introduction: For Africans living in the United States, the extent to which stress-induced physiologic dysfunction varies by reason for immigration is unknown. Allostatic load score (ALS) is used to assess physiologic stress. ALS has biomarkers in 3 domains: cardiovascular, metabolic and immune. However, there are many ALS equations and the number of biomarkers in each domain varies. In the cardiac domain of ALS, equations use either triglyceride (TG) or high density lipoprotein (HDL). A specific problem for African descent populations is that HDL may be superior to TG as a marker of insulin resistance and cardiac risk. Objective: Our primary goal in 95 African immigrants (71% male, age 42±10, (mean±SD), range 22-62y) was to determine if ALS varied by reason for immigration. Our secondary goal was to evaluate 4 different ALS equations to determine if there was a difference in ALS performance depending on whether HDL or TG was the included biomarker. Methods and Results: For all 4 equations, the reasons for immigration from lowest to highest ALS were: family reunification, lottery for self and immediate family, marriage, asylum, work and study (Figure 1). As the first 3 reasons for immigration promoted family unity, they were grouped together (Group 1). The Africans who came for other reasons were grouped together (Group 2). Equations 1 and 2 included HDL as a biomarker. For these equations, ALS in Group 1 vs. Group 2 were: 1.7±1.3 vs. 2.8±1.7, P <0.01 and 1.4±1.2 vs. 2.3±1.7, P <0.01 resp. Equations 3 and 4 used TG. For these two equations ALS in Group 1 vs. Group 2 were: 2.2±1.6 vs. 2.9±2.0, P =0.08 and 2.1±1.4 vs. 2.9±2.0, P =0.03 resp. In Equations 1 and 2, when HDL was replaced by TG, the significant difference in ALS between groups declined. In Equations 3 and 4, when TG was replaced by HDL, the significant difference in ALS between groups increased. Conclusions: ALS varies by reason for immigration and is easier to detect in ALS equations which use HDL rather than TG.


2021 ◽  
Vol 9 (4) ◽  
pp. 65-68
Author(s):  
Noor Muhammad Marwat ◽  
Shah Khalid ◽  
Pir Muhammad Abdul Aziz Shah ◽  
Fayaz ul Hasnain ◽  
Rashid Naeem Khan

Purpose: The purpose of this study to find out the impacts of a 6-weeks supervised aerobic exercise protocol on High-Density Lipoprotein among adults aged between 25-35 years was evaluated. Methodology: Researchers through a non-probability sampling procedure selected thirty (30) volunteers ranging from 25-35 untrained adults from Lakki Marwat. Researchers used a six-week aerobic exercise protocol to collect relevant information from the targeted dependent variables. The CHOD PAP method was used to measure the High-Density Lipoprotein (HDL) of adults. All the data collected from pre-and post- (HDL) tests were recorded in numerical form and analyzed by using a t-test. Main Findings: The results of the study indicate that the six-week aerobic exercise protocol program had no significant difference on pre-and post-intervention quantities of HDL of Experimental Group which is (P>0.05). The implication of the Study: The aerobic exercise programs used for this particular study may help adults to improve and maintain their health status, proper social stature, and lipid profile. An increase in the HLD is ideal as this increase in the HDL helps to prevent cardiovascular diseases particularly the heart valves from stroke and cardiac arrest and academic achievements of the participants. Novelty: So far, no particular research has been conducted on the social implications of exercise effects on adults. Future researchers may work on the social habits of physically fit adults and their input to society.


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