Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S]
Endogenous labeling with stable isotopes is used to study the metabolism of proteins in vivo. However, traditional detection methods such as GC/MS cannot measure tracer enrichment in multiple proteins simultaneously, and multiple reaction monitoring MS cannot measure precisely the low tracer enrichm...
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| Format: | Article |
| Language: | English |
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Elsevier
2016-04-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520354262 |
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| author | Sasha A. Singh Allison B. Andraski Brett Pieper Wilson Goh Carlos O. Mendivil Frank M. Sacks Masanori Aikawa |
| author_facet | Sasha A. Singh Allison B. Andraski Brett Pieper Wilson Goh Carlos O. Mendivil Frank M. Sacks Masanori Aikawa |
| author_sort | Sasha A. Singh |
| collection | DOAJ |
| description | Endogenous labeling with stable isotopes is used to study the metabolism of proteins in vivo. However, traditional detection methods such as GC/MS cannot measure tracer enrichment in multiple proteins simultaneously, and multiple reaction monitoring MS cannot measure precisely the low tracer enrichment in slowly turning-over proteins as in HDL. We exploited the versatility of the high-resolution/accurate mass (HR/AM) quadrupole Orbitrap for proteomic analysis of five HDL sizes. We identified 58 proteins in HDL that were shared among three humans and that were organized into five subproteomes according to HDL size. For seven of these proteins, apoA-I, apoA-II, apoA-IV, apoC-III, apoD, apoE, and apoM, we performed parallel reaction monitoring (PRM) to measure trideuterated leucine tracer enrichment between 0.03 to 1.0% in vivo, as required to study their metabolism. The results were suitable for multicompartmental modeling in all except apoD. These apolipoproteins in each HDL size mainly originated directly from the source compartment, presumably the liver and intestine. Flux of apolipoproteins from smaller to larger HDL or the reverse contributed only slightly to apolipoprotein metabolism. These novel findings on HDL apolipoprotein metabolism demonstrate the analytical breadth and scope of the HR/AM-PRM technology to perform metabolic research. |
| first_indexed | 2024-12-20T11:20:12Z |
| format | Article |
| id | doaj.art-845a42f232bc4a5699765ac5050d25c9 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-20T11:20:12Z |
| publishDate | 2016-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-845a42f232bc4a5699765ac5050d25c92022-12-21T19:42:31ZengElsevierJournal of Lipid Research0022-22752016-04-01574714728Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S]Sasha A. Singh0Allison B. Andraski1Brett Pieper2Wilson Goh3Carlos O. Mendivil4Frank M. Sacks5Masanori Aikawa6Center for Interdisciplinary Cardiovascular Sciences, Cardiovascular Division Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MADepartment of Nutrition, Harvard T. H. Chan School of Public Health, Boston, MACenter for Interdisciplinary Cardiovascular Sciences, Cardiovascular Division Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MACenter for Interdisciplinary Cardiovascular Sciences, Cardiovascular Division Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MASchool of Medicine, Universidad de los Andes, Bogota, ColombiaTo whom correspondence should be addressed. (M.A.); (F.M.S.); Department of Nutrition, Harvard T. H. Chan School of Public Health, Boston, MA; Channing Division of Network Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MATo whom correspondence should be addressed. (M.A.); (F.M.S.); Center for Interdisciplinary Cardiovascular Sciences, Cardiovascular Division Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA; Channing Division of Network Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MAEndogenous labeling with stable isotopes is used to study the metabolism of proteins in vivo. However, traditional detection methods such as GC/MS cannot measure tracer enrichment in multiple proteins simultaneously, and multiple reaction monitoring MS cannot measure precisely the low tracer enrichment in slowly turning-over proteins as in HDL. We exploited the versatility of the high-resolution/accurate mass (HR/AM) quadrupole Orbitrap for proteomic analysis of five HDL sizes. We identified 58 proteins in HDL that were shared among three humans and that were organized into five subproteomes according to HDL size. For seven of these proteins, apoA-I, apoA-II, apoA-IV, apoC-III, apoD, apoE, and apoM, we performed parallel reaction monitoring (PRM) to measure trideuterated leucine tracer enrichment between 0.03 to 1.0% in vivo, as required to study their metabolism. The results were suitable for multicompartmental modeling in all except apoD. These apolipoproteins in each HDL size mainly originated directly from the source compartment, presumably the liver and intestine. Flux of apolipoproteins from smaller to larger HDL or the reverse contributed only slightly to apolipoprotein metabolism. These novel findings on HDL apolipoprotein metabolism demonstrate the analytical breadth and scope of the HR/AM-PRM technology to perform metabolic research.http://www.sciencedirect.com/science/article/pii/S0022227520354262coronary heart diseasesize fractionationmass spectrometrycompartmental modelingmetabolism |
| spellingShingle | Sasha A. Singh Allison B. Andraski Brett Pieper Wilson Goh Carlos O. Mendivil Frank M. Sacks Masanori Aikawa Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] Journal of Lipid Research coronary heart disease size fractionation mass spectrometry compartmental modeling metabolism |
| title | Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] |
| title_full | Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] |
| title_fullStr | Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] |
| title_full_unstemmed | Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] |
| title_short | Multiple apolipoprotein kinetics measured in human HDL by high-resolution/accurate mass parallel reaction monitoring[S] |
| title_sort | multiple apolipoprotein kinetics measured in human hdl by high resolution accurate mass parallel reaction monitoring s |
| topic | coronary heart disease size fractionation mass spectrometry compartmental modeling metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520354262 |
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