Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver.
Nascent high density lipoprotein (HDL) (1.063 less than d less than 1.21 g/ml) was isolated from recirculating rat liver perfusates and separated by heparin-Sepharose chromatography into a nonretained fraction (NR) and a fraction (R) that eluted with 0.5 M NaCl. Fractions NR and R contained 70% and...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
1989-07-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520383000 |
| _version_ | 1819058224258613248 |
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| author | K E Winkler J B Marsh |
| author_facet | K E Winkler J B Marsh |
| author_sort | K E Winkler |
| collection | DOAJ |
| description | Nascent high density lipoprotein (HDL) (1.063 less than d less than 1.21 g/ml) was isolated from recirculating rat liver perfusates and separated by heparin-Sepharose chromatography into a nonretained fraction (NR) and a fraction (R) that eluted with 0.5 M NaCl. Fractions NR and R contained 70% and 30% of the nascent HDL protein, respectively. ApoB-containing particles were removed from fraction R by chromatography on concanavalin A. The protein composition of fractions NR and R was 40% and 29%, respectively. Fraction NR contained 25% apoA-I, 11% apoA-IV, 24% apoE, and 38% apoC. Fraction R contained primarily apoE (81% of total protein). The lipid composition of NR and R, respectively, was: triglyceride 44% and 26%, phospholipid 41% and 57%, cholesterol 8% and 13%, and cholesteryl ester 7% and 4%. Fractions NR and R had molecular weights of 400,000 and 860,000, respectively, as calculated from the Stokes radius. Negative staining electron microscopy indicated that both fractions consisted mainly of spherical particles (260-280 A) but some stacked disks were seen in fraction R. Livers perfused by the single-pass technique produced fractions NR and R in the same ratio as livers perfused by recirculation. The apolipoprotein compositions were similar to those in the recirculating perfusion; however, both fractions NR and R had more triglyceride (greater than 50% of total lipid). An HDL fraction was also isolated from liver perfusates by a combination of molecular sieve and heparin-Sepharose affinity chromatography. This HDL contained triglyceride but no apoB, indicating that triglyceride-rich HDL particles are not an artifact of ultracentrifugation.(ABSTRACT TRUNCATED AT 250 WORDS) |
| first_indexed | 2024-12-21T13:51:48Z |
| format | Article |
| id | doaj.art-bba8d9c234c14da09c9be4ba3b1f06e9 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-21T13:51:48Z |
| publishDate | 1989-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-bba8d9c234c14da09c9be4ba3b1f06e92022-12-21T19:01:40ZengElsevierJournal of Lipid Research0022-22751989-07-01307979987Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver.K E Winkler0J B Marsh1Department of Physiology and Biochemistry, Medical College of Pennsylvania, Philadelphia 19129.Department of Physiology and Biochemistry, Medical College of Pennsylvania, Philadelphia 19129.Nascent high density lipoprotein (HDL) (1.063 less than d less than 1.21 g/ml) was isolated from recirculating rat liver perfusates and separated by heparin-Sepharose chromatography into a nonretained fraction (NR) and a fraction (R) that eluted with 0.5 M NaCl. Fractions NR and R contained 70% and 30% of the nascent HDL protein, respectively. ApoB-containing particles were removed from fraction R by chromatography on concanavalin A. The protein composition of fractions NR and R was 40% and 29%, respectively. Fraction NR contained 25% apoA-I, 11% apoA-IV, 24% apoE, and 38% apoC. Fraction R contained primarily apoE (81% of total protein). The lipid composition of NR and R, respectively, was: triglyceride 44% and 26%, phospholipid 41% and 57%, cholesterol 8% and 13%, and cholesteryl ester 7% and 4%. Fractions NR and R had molecular weights of 400,000 and 860,000, respectively, as calculated from the Stokes radius. Negative staining electron microscopy indicated that both fractions consisted mainly of spherical particles (260-280 A) but some stacked disks were seen in fraction R. Livers perfused by the single-pass technique produced fractions NR and R in the same ratio as livers perfused by recirculation. The apolipoprotein compositions were similar to those in the recirculating perfusion; however, both fractions NR and R had more triglyceride (greater than 50% of total lipid). An HDL fraction was also isolated from liver perfusates by a combination of molecular sieve and heparin-Sepharose affinity chromatography. This HDL contained triglyceride but no apoB, indicating that triglyceride-rich HDL particles are not an artifact of ultracentrifugation.(ABSTRACT TRUNCATED AT 250 WORDS)http://www.sciencedirect.com/science/article/pii/S0022227520383000 |
| spellingShingle | K E Winkler J B Marsh Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. Journal of Lipid Research |
| title | Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. |
| title_full | Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. |
| title_fullStr | Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. |
| title_full_unstemmed | Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. |
| title_short | Characterization of nascent high density lipoprotein subfractions from perfusates of rat liver. |
| title_sort | characterization of nascent high density lipoprotein subfractions from perfusates of rat liver |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520383000 |
| work_keys_str_mv | AT kewinkler characterizationofnascenthighdensitylipoproteinsubfractionsfromperfusatesofratliver AT jbmarsh characterizationofnascenthighdensitylipoproteinsubfractionsfromperfusatesofratliver |