The Ultracentrifugal Characterization and Isolation of Human Blood Lipids and Lipoproteins, with Applications to the Study of Atherosclerosis.

Lindgren, Frank T.; Elliott, Harold; Gofman, John W.

doi:10.1021/j150484a010

Cited by 216 publications

(57 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Most ofthe reported studies (Lindgren et al, 1951(Lindgren et al, , 1969Adams & Schumaker, 1969) on the molecularweight determination of LD lipoprotein preparations were based on the assumption that these molecules are anhydrous spheres; the measured parameters were flotation coefficients and hydrated densities. Lindgren et al (1969) found a molecular weight of 2.12 x 106 for LD lipoprotein from men and 2.36 x 106 for women, whereas Adams & Schumaker (1969) found values of 2.1 x 106-2.6 x 106.…”

Section: Discussionmentioning

confidence: 99%

Physicochemical properties of low-density lipoproteins of normal human plasma. Evidence for the occurrence of lipoprotein B in associated and free forms

Lee

Alaupovic

1974

Biochemical Journal

View full text Add to dashboard Cite

1. Low-density (d 1.006-1.063 g/ml) lipoproteins from normal human plasma were separated by differential preparative ultracentrifugation into six subfractions. Each lowdensity (LD)lipoprotein subfractioncontained lipoprotein B as the major andlipoproteins A and C as the minor lipoprotein families. 2. Three lipoprotein B subfractions (LP-B), LP-B-III (d 1.019-1 .030g/ml), LP-B-IV (d 1.030-1.040g/ml) and LP-B-V (d 1.040-1.053 g/ ml) were prepared from the corresponding LD lipoprotein subfractions by immunoprecipitating small amounts oflipoproteins A and C. 3. Determination of hydrodynamic properties indicated that LD lipoproteins consisted of three molecular segments characterized by a stepwise change in the molecular weight: LDL-I and LDL-II subfractions (d 1.006-1.019g/ml) with an average mol.wt. of 4.75 x 106, LDL-III (d 1.019-1.030g/ml) with a mol.wt. of 3.99 x 106, and LDL-IV, LDL-V and LDL-VI (d 1.030-1.053g/ml) with a mol.wt. of 2.85 x 106. 4. All three lipoprotein B subfractions had an average mol.wt. of 3.16 x 106. 5. The LDL-I and LDL-II subfractions consisted of lipoprotein B and lipoprotein C families which were present in the form of an association complex. This was isolated from serum by immunoprecipitation with antibodies to lipoprotein B. The complex had a mol.wt. of 4.35 x 106. 6. The results indicate a fundamental difference between the LD lipoprotein subfractions with d 1.006-1.019g/ml and those subfractions with d 1.030-1.063g/ml. In the former, lipoprotein B occurs as a part of an association complex, whereas in the latter it occurs as a separate entity.Human plasma low-density lipoproteins consist of two macromolecular distributions characterized by Sf 0-12 and Sf 12-20 (Oncley, 1963;Ewing et al., 1965;Nichols, 1967). The heterogeneity of both subclasses in regard to particle size, hydrated density and molecular weight has been attributed to discrete changes in the lipid and protein content of the lipoprotein molecules (Oncley etal., 1957;Shore & Shore, 1962;Nichols, 1967;Dearborn & Wetlaufer, 1969

show abstract

Section: Discussionmentioning

confidence: 99%

Physicochemical properties of low-density lipoproteins of normal human plasma. Evidence for the occurrence of lipoprotein B in associated and free forms

Lee

Alaupovic

1974

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…In 1954, John Gofman and colleagues reported that analytic ultracentrifuge measurements of high-density lipoproteins (HDL) in blood revealed two particle subclasses, HDL2 and HDL3, and that the concentrations of the more buoyant particles (HDL2) were 50% higher in women than men ( 1,2 ). In 1966, they reported that 38 men who developed coronary heart disease (CHD) had HDL mass for HDL2 than HDL3 ( 17 ).…”

mentioning

confidence: 99%

Fifty-three year follow-up of coronary heart disease versus HDL2 and other lipoproteins in Gofman's Livermore Cohort

Williams

2012

Journal of Lipid Research

View full text Add to dashboard Cite

This article is available online at http://www.jlr.org concentrations that were 32% lower for HDL2 and 8% lower for HDL3 (the less buoyant particles) compared with those who did not develop CHD ( 3 ).Gofman's initial observation gave rise to studies by others on the potential clinical utility of HDL2 and HDL3 measurements. Rather than employing Gofman's original methodology, they measured HDL2-and HDL3-cholesterol by precipitation and HDL size classes by nuclear magnetic resonance (NMR) or, more rarely, ultracentrifugation. Many studies showed that high HDL2 was associated with lower CHD risk ( 4-10 ); however, the independent effects of HDL2 and HDL3 on CHD risk remained inconclusive ( 5, 7-10 ). Currently, the National Cholesterol Education Program Adult Treatment Panel (ATP-III) guidelines ( 11 ) state that "although small studies suggest greater predictive power of one or another HDL component, their superiority over HDL-cholesterol has not been demonstrated in large, prospective studies. Consequently, ATP III does not recommend the routine measurement of HDL subspecies in CHD risk assessment." However, the clinical utility of HDL subclasses may be underappreciated in part due to the limitations of these alternative methods to adequately characterize HDL heterogeneity ( 12-16 ).Between 1954 and 1956, John Gofman created a cohort of 1,905 men in whom lipoproteins were measured by analytic ultracentrifugation ( 3 ). Our rediscovery of Gofman's original data enabled us to assess the relationships of lipoprotein mass concentrations to CHD during 29-year follow-up ( 17 ). Those analyses showed that i ) the lowest quartile of HDL2-mass increased the men's risks of fatal plus nonfatal CHD by 38% and their risks of premature CHD (age р 65 years) by 61% when adjusted for traditional risk factors, ii ) the HDL2-CHD risk relationship remained signifi cant when adjusted for HDL3, and iii ) the risk reduction per mg/dl of HDL-mass was signifi cantly greater Abstract To assess the relationships of lipoprotein mass concentrations to all-cause and coronary heart disease (CHD) mortality, we analyzed the prospective 53-year follow-up of 1,905 men measured for lipoprotein mass concentrations by analytic ultracentrifugation between 1954 and 1957. Cause of death was determined from medical records and death certifi cates before 1979 and from National Death Index death diagnoses thereafter. Of the 1,329 men (69.8%) who died through 2008 , CHD was listed as a contributing cause of death for 409 men, including 113 deaths from premature CHD (age р 65 years). When adjusted for age, the risk associated with the lowest HDL2 quartile increased 22% for all-cause ( P = 0.001), 63% for total CHD ( P < 10 ؊ 5 ), and 117% for premature CHD mortality ( P = 0.0001). When adjusted for standard risk factors (age, total cholesterol, blood pressure, BMI, smoking) and the lowest HDL3 quartile, the corresponding risk increases were 14% ( P = 0.05), 38% ( P = 0.004), and 62% ( P = 0.02), respectively. Men with HDL3 р 25 th percentile had 28% greater ...

show abstract

“…The other possibility is that large molecules containing these lipids have the observed distribution because of their density. Lindgren, Elliott, and Gofman have published in graphic form the results of similarly prolonged centrifugation of untreated serum (17). Our graph in Figure 3 shows a number of similarities to and differences from theirs.…”

Section: Resultsmentioning

confidence: 58%

“…Our observation that the specific gravity of centrifugate of our top sample is approximately that of water can-only be explained by the presence of lipid aggregates having a lower specific gravity than that of the medium in which they are suspended. Lindgren, Elliott and Gofman have reported that chylomicrons and other lipid particles have a specific gravity of less than 1.006 (17). Obviously the two ends of the column represent a special condition where lipid complexes may be stopped while moving toward their own density levels.…”

Section: Resultsmentioning

confidence: 99%

The Study of Serum Proteins and Lipids With the Aid of the Quantity Ultracentrifuge. I. Procedure and Principal Features of the Centrifugate of Untreated Normal Serum as Determined by Quantitative Analysis of Samples From Ten Levels 12

Turner¹,

Snavely²,

Goldwater³

et al. 1951

J. Clin. Invest.

View full text Add to dashboard Cite

The studies described in this paper and some which follow have as an objective a better description of serum lipids. The present paper represents the first detailed report of work begun early in 1948, some features of which have been published in abstract (1)(2)(3)(4)(5). Initially our purpose was to extend the observations of Recant, Chargaff and Hanger (6) on the influence of serum proteins upon the cephalin-cholesterol flocculation and the thymol turbidity tests. We included quantitative analyses of the centrifugate for lipids as well as for proteins. It soon became apparent that we were using a procedure which was useful in the study of the structure and function of lipid complexes.As shown in the review by Edsall (7) there has long been evidence that a large proportion of the lipids of the serum occur not as small molecules, but as lipid aggregates, or lipoproteins, of varying complexity. The use of the ultracentrifuge, both quantitative and analytical, has a well established place in the study of serum lipids. Important contributions have been made by McFarlane (8,9), Pedersen (10, 11), the Harvard group (12) ment of rate of flotation when suspended in a standard medium, a technic which we have not used. Principal emphasis in published studies by other workers has been on centrifugation of treated serum. In the present paper we deal exclusively with centrifugation of untreated whole serum. At present it does not seem profitable to attempt a detailed comparison between the results obtained by our procedure on the one hand and those obtained by the system of analysis introduced by Cohn and his associates (21), and the results of the flotation procedures of others. When our studies of diseased states and normal individuals under dietary stress have been presented, such comparisons will be in order. Some of the similarities of our contributions to those of others will be mentioned below in the discussion.The perfect procedure for the study of serum proteins and lipids in the initial step would give complete separation of the different lipid and protein complexes, each entity in pure state with structure unchanged by the procedure effecting separation. We doubt if the procedure described in this communication separates any single lipid or protein complex in pure form; nevertheless, the concentration of complexes at certain levels in the centrifugate is of such a degree as to give new information concerning the structure of the complexes at that level and their concentration in the serum under study. The lipid distribution curves are reproducible within satisfactory limits with normal sera, and are significantly different in certain diseased states, especially in acute virus hepatitis, as we have already reported briefly (3) and will report later in detail. We believe our procedure has three main advantages: The manipulation of serum which effects redistribution of pro-1071 TURNER, SNAVELY, GOLDWATER, RANDOLPH, SPRAGUE, AND UNGLAUB teins and lipids is gentle and involves minimal risk of damage to complex molecul...

show abstract

The Ultracentrifugal Characterization and Isolation of Human Blood Lipids and Lipoproteins, with Applications to the Study of Atherosclerosis.

Cited by 216 publications

References 0 publications

Physicochemical properties of low-density lipoproteins of normal human plasma. Evidence for the occurrence of lipoprotein B in associated and free forms

Physicochemical properties of low-density lipoproteins of normal human plasma. Evidence for the occurrence of lipoprotein B in associated and free forms

Fifty-three year follow-up of coronary heart disease versus HDL2 and other lipoproteins in Gofman's Livermore Cohort

The Study of Serum Proteins and Lipids With the Aid of the Quantity Ultracentrifuge. I. Procedure and Principal Features of the Centrifugate of Untreated Normal Serum as Determined by Quantitative Analysis of Samples From Ten Levels 12

Contact Info

Product

Resources

About