Characterization of human high-density lipoproteins by gradient gel electrophoresis

Blanche, Patricia J.; Gong, Enhao; Forte, Trudy M.; Nichols, Alex V.

doi:10.1016/0005-2760(81)90253-8

Cited by 577 publications

(269 citation statements)

References 9 publications

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“…14 New techniques for HDL separation have identified a number of discrete subpopulations of particles of varying size and composition. 15 - 17 The present study was undertaken to further investigate the relations of individual HDL subclasses as determined by gradient gel electrophoresis to the progression of premature coronary atherosclerosis as assessed by repeated angiographies. Young survivors of myocardial infarction (MI) who had routinely undergone coronary angiography after the acute event were reinvestigated by HDL subfractionation, determination of the lipid contents of the major lipoprotein classes, and a second coronary angiography approximately 5 years after the baseline examination.…”

Section: Ost Prospective Epidemiological Studiesmentioning

confidence: 99%

“…HDL subclasses were determined by a modification of the gradient gel electrophoresis method described by Blanche et al 15 In principle, HDL of d=1.070-1.21 kg/1 was first separated from plasma by two consecutive runs in a 50.3 Ti fixed-angle rotor (model L 8-80 ultracentrifuge Beckman, Palo Alto, Calif.). In the first step of the HDL preparation, 1 ml d= 1.3106 kg/1 NaBr solution was added to 4 ml plasma in a Belltop tube, Beckman (final d=1.070 kg/1), and ultracentrifugation was then performed for 20 hours (50,000 rpm, 179,200g, 1°C).…”

Section: High Density Lipoprotein Gradient Gel Electrophoresismentioning

confidence: 99%

“…To estimate the protein concentration of each HDL subclass, the protein concentration of plasma HDL was multiplied with the relative HDL subclass areas from the scanning plot. Partcle diameter (d) ranges for the HDL subclasses were defined according to Blanche et al 15 as follows: HDL 2b , 9.71 <d< 12.9 nm; HDL^, 8.77<d<9.71 nm; HDL3,, 8.17<d< 8.77 nm; HDL 3b , 7.76<d<8.17 nm; and HDL^ 7.21<d<7.76 nm. The location of these limits on the scanning curves was derived from the relative migration of the protein standards with their known Stokes' diameters in the gels.…”

Section: High Density Lipoprotein Gradient Gel Electrophoresismentioning

confidence: 99%

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High density lipoproteins and coronary atherosclerosis. A strong inverse relation with the largest particles is confined to normotriglyceridemic patients.

Johansson¹,

Carlson²,

Landou³

et al. 1991

Arterioscler Thromb

239

117

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The relations of high density lipoprotein (HDL) subclasses to severity and rate of progression of coronary atherosclerosis were investigated in 60 men who had survived a myocardial infarction before the age of 45 years and who had subsequently undergone two coronary angjographies, with an intervening time interval of 4-7 years between angiographies. Five HDL subclasses with different particle sizes were determined by gradient gel electrophoresis, and the major serum lipoprotein classes were separated by preparative ultracentrifugation in connection with the second angiography. Highly significant inverse correlations were found between the plasma levels of the largest HDL particles, the HDLa, subclass, and both disease severity as observed on the second coronary angiogram (r=-0.53, p<0.001) and progression of coronary lesions between angiographies (r=-038, /?<0.01). Grouping the patients according to the presence or absence of very low density lipoprotein (VLDL) triglyceride elevation revealed striking differences in the relations of HDL subspecies to coronary atherosclerosis between normotriglyceridemic and hvpertriglyceridemic subjects. There were strong inverse correlations between the plasma HDL^ concentration and both severity of lesions (r=-0.72,p<0.001) and rate of lesion progression (r=-0.58, p<0.01) in the normotriglyceridemic patients, whereas this relation was absent in subjects with hypertriglyceridemia. Although it was retrospective in design, affected by an inherent selection bias due to unavoidable dropouts and exclusions, and limited to young, male, postinfarction patients, the study suggests that the inverse relation between HDL level and coronary heart disease or coronary atherosclerosis seen in previous epidemiological and angiographlc studies is accounted for by the largest HDL particles, the HDLy, subclass. Furthermore, the hvpertriglyceridemic state appears to markedly influence the relations between plasma concentrations of HDL subclasses and coronary atherosclerosis. Prospective studies of unselected patients are needed to corroborate these findings. (Arteriosclerosis and Thrombosis 1991;11:174-182) M ost prospective epidemiological studies have shown an independent inverse relation between high density lipoprotein (HDL) cholesterol concentration and risk of development of coronary heart disease (CHD).

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Section: Ost Prospective Epidemiological Studiesmentioning

confidence: 99%

Section: High Density Lipoprotein Gradient Gel Electrophoresismentioning

confidence: 99%

Section: High Density Lipoprotein Gradient Gel Electrophoresismentioning

confidence: 99%

See 1 more Smart Citation

High density lipoproteins and coronary atherosclerosis. A strong inverse relation with the largest particles is confined to normotriglyceridemic patients.

Johansson¹,

Carlson²,

Landou³

et al. 1991

Arterioscler Thromb

239

117

View full text Add to dashboard Cite

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“…These subclasses are defined by their estimated particle diameters: HDL3c (approximately 7.2-7.8 om), HDL3b (7.8-8.2 nm), HDL3a (8:2-8.8 om), HDL2a (8.8-9.7) and HDL2b (9.7-12.9 om) [Blanche et al, 1981]. HDL particles may contain both apoA-I and apoA-II or apoA-I without apo-AII [Cheung et al, 1984].…”

mentioning

confidence: 99%

“…Plasma was ultracentrifuged at d<1.006 g/ml, cholesterol determined in the infranant, and LDL-cholesterol was_calculated from the formula by Friedewald et al [1972] . Plasma apolipoprotein A-I concentrations were determined by radialimmunodiffusion [Cheung et al, 1977] 0 Electrophoresis of HDL in the ultracentrifuged d ~ 1.20 g/ml fraction was performed on Pharmacia Electrophoresis Apparatus (GE 4-II) using slab gradient gels (PAA 4/30, Pharmacia, Piscataway, NJ) as described by Blanche et al [1981].…”

mentioning

confidence: 99%

Lipoprotein subclasses in genetic studies: The Berkeley data set

Krauss

Williams

Blanche

et al. 1993

Genetic Epidemiology

Self Cite

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In conjunction with a study examining the inheritance of LDL subclass patterns in a healthy population, measurements of lipids, lipoproteins, and lipoprotein subclasses were performed in 301 individuals in 27 kindreds. Questionnaires were used to obtain information on use of medications, hormones, cigarettes, and alcohol. Laboratory data from this study (the Berkeley data set) include measurements of LDL and HDL size subclasses by nondenaturing gradient gel electrophoresis, and measurement of apolipoprotein A‐I by radial immunodiffusion. © 1993 Wiley‐Liss, Inc.

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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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Characterization of human high-density lipoproteins by gradient gel electrophoresis

Cited by 577 publications

References 9 publications

High density lipoproteins and coronary atherosclerosis. A strong inverse relation with the largest particles is confined to normotriglyceridemic patients.

High density lipoproteins and coronary atherosclerosis. A strong inverse relation with the largest particles is confined to normotriglyceridemic patients.

Lipoprotein subclasses in genetic studies: The Berkeley data set

Chromatography of Membrane Proteins and Lipoproteins

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