The determination of specific radioactivity of proteins eluted intact from polyacrylamide gels, utilizing a fluorescamine assay

Kilgore, Laura L.; Rogers, Julie L.; Patterson, Bruce W.; Miller, Nancy Houston; Fisher, Waldo R.

doi:10.1016/0003-2697(85)90335-5

Cited by 9 publications

(14 citation statements)

References 7 publications

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“…The proteins were then isolated in the same Tris buffer but with 6 M urea using, respectively, either Sephacryl S-200 or Sepharose CL-6B columns, and the purity ofthe proteins was determined by polyacrylamide gel electrophoresis (18). Apolipoproteins from VLDL and HDL were also purified directly from polyacrylamide gels by electrophoretic elution as recently described (19 specifically.…”

Section: Methodsmentioning

confidence: 99%

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Kilgore¹,

Patterson²,

Parenti³

et al. 1985

J. Clin. Invest.

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An antibodylike paraprotein has been isolated from a patient with multiple myeloma and autoimmune hyperlipoproteinemia.The paraprotein bound to apolipoprotein B (apo B)-containing lipoproteins that formed macromolecular aggregates, and globules thought to be aggregated complexes of lipoproteins and reactive immunoglobulins were observed circulating within the retinal blood vessels of this patient. This binding specificity permitted purification of the paraprotein from both the agglutinated immune complexes and from the plasma.The protein is an IgA, K-immunoglobulin which exists primarily in a polymeric state. Capillary immunoprecipitation demonstrated reactivity with very low density lipoproteins (VLDL) and low density proteins (LDL), but not with high density lipoproteins (HDL). Delipidated apo B and apo E, but not apo A or apo C, formed precipitates with this immunoglobulin. In using a radioimmunoassay format, the affinity of the immunoglobulin was greatest for VLDL

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Section: Methodsmentioning

confidence: 99%

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Kilgore¹,

Patterson²,

Parenti³

et al. 1985

J. Clin. Invest.

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“…Lipoproteins formed by this pathway initially appear in plasma as either large VLDL, L (13,12) or as smaller IDL, L (16,12). Figure 2 shows the model-generated fit to VLDL and the simulations resulting when alternately L (13,12) and L (18,9) have been set to zero, thus demonstrating the requirement for both pathways in fitting these data. While large VLDL-apoB that enters C( 18) is rapidly converted to IDL, that entering C( 13) is metabolized along a three-compartment delipidation chain, as previously described, enroute to IDL (12).…”

Section: Apob Modelmentioning

confidence: 91%

“…Finally, LDL was isolated at density 1.06 g/ml for 20 h. Recovered lipoproteins were frozen in 20% sucrose pending isolation of apoB by preparative, acrylamide gel electrophoresis in an SDS buffer. The recovered apoB was quantitated by a fluorescamine protein assay, and specific radioactivity was determined by scintillation counting, as described previously (9).…”

Section: Fractionation Of Plasma Lipoproteins and Measurement Of Apob Specific Radioactivitymentioning

confidence: 99%

“…Metabolic channeling is an important feature of apoB physiology (16,17). Approximately one-quarter of apoB is secreted as large VLDL, L (18,9), one-third as IDL, L (27,9), and 5% as LDL, L (30,9), in what constitutes a fast path for the channeling of apoB to LDL. An additional 40% appears in plasma after a delay, L (12,9), which is thought to result from the time required in the hepatocyte to package additional lipid in an enriched fraction of VLDL particles (17).…”

Section: Apob Modelmentioning

confidence: 99%

“…Approximately one-quarter of apoB is secreted as large VLDL, L (18,9), one-third as IDL, L (27,9), and 5% as LDL, L (30,9), in what constitutes a fast path for the channeling of apoB to LDL. An additional 40% appears in plasma after a delay, L (12,9), which is thought to result from the time required in the hepatocyte to package additional lipid in an enriched fraction of VLDL particles (17). Lipoproteins formed by this pathway initially appear in plasma as either large VLDL, L (13,12) or as smaller IDL, L (16,12).…”

Section: Apob Modelmentioning

confidence: 99%

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Apolipoprotein B metabolism in hypertriglyceridemic diabetic patients administered either a fish oil- or vegetable oil-enriched diet

Fisher

Zech

Stacpoole

1998

Journal of Lipid Research

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The effect on apolipoprotein B kinetics of a diet enriched in either fish oil or safflower oil was investigated in five hypertriglyceridemic (HTG), non-insulin-dependent diabetic subjects. The fish oil diet decreased plasma triglycerides and VLDL-apoB but increased LDL-apoB and LDL-cholesterol. Total plasma apoB concentration did not change, nor did the increased VLDL-apoB secretion present in these HTG subjects, which, accompanied by impaired lipolysis, accounted for their elevated VLDL. The fish oil-induced fall in VLDL resulted from a decrease in secretion without a change in residence time. The IDL fraction, which also contained small VLDL, was the primary site for the secretion of apoB particles in the HTG subjects. On the fish oil diet there was a further, compensatory increase in the secretion of these lipoproteins such that the transport of apoB in IDL remained the same, as did its mass. In the HTG subjects the major portion of IDL lipoproteins was catabolized, with LDL-apoB production comprising the lesser quantity. On the fish oil diet, a shift in the channeling of the lipoprotein output from IDL resulted in a decrease in the catabolic pathway and an increase in conversion to LDL. As the residence time of LDL did not change, this increased input gave rise to the larger mass of LDL-apoB seen in these hypertriglyceridemic subjects when receiving a fish oil diet.-Fisher, W. R., L. A. Zech, and P. W. Stacpoole. Apolipoprotein B metabolism in hypertriglyceridemic diabetic patients administered either a fish oil-or vegetable oilenriched diet.

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[11] Isolation and characterization of apolipoprotein B-100

Fisher¹,

Schumaker²

1986

Methods in Enzymology

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The determination of specific radioactivity of proteins eluted intact from polyacrylamide gels, utilizing a fluorescamine assay

Cited by 9 publications

References 7 publications

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Immune complex hyperlipidemia induced by an apolipoprotein-reactive immunoglobulin A paraprotein from a patient with multiple myeloma. Characterization of this immunoglobulin.

Apolipoprotein B metabolism in hypertriglyceridemic diabetic patients administered either a fish oil- or vegetable oil-enriched diet

[11] Isolation and characterization of apolipoprotein B-100

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