Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles
The partitioning of apolipoprotein A-I (apoA-I) molecules in plasma between HDL-bound and -unbound states is an integral part of HDL metabolism. We used the surface plasmon resonance (SPR) technique to monitor in real time the reversible binding of apoA-I to HDL. Biotinylated human HDL2 and HDL3 wer...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2010-03-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520305186 |
| _version_ | 1818732466908692480 |
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| author | Sissel Lund-Katz David Nguyen Padmaja Dhanasekaran Momoe Kono Margaret Nickel Hiroyuki Saito Michael C. Phillips |
| author_facet | Sissel Lund-Katz David Nguyen Padmaja Dhanasekaran Momoe Kono Margaret Nickel Hiroyuki Saito Michael C. Phillips |
| author_sort | Sissel Lund-Katz |
| collection | DOAJ |
| description | The partitioning of apolipoprotein A-I (apoA-I) molecules in plasma between HDL-bound and -unbound states is an integral part of HDL metabolism. We used the surface plasmon resonance (SPR) technique to monitor in real time the reversible binding of apoA-I to HDL. Biotinylated human HDL2 and HDL3 were immobilized on a streptavidin-coated SPR sensor chip, and apoA-I solutions at different concentrations were flowed across the surface. The wild-type (WT) human and mouse apoA-I/HDL interaction involves a two-step process; apoA-I initially binds to HDL with fast association and dissociation rates, followed by a step exhibiting slower kinetics. The isolated N-terminal helix bundle domains of human and mouse apoA-I also exhibit a two-step binding process, consistent with the second slower step involving opening of the helix bundle domain. The results of fluorescence experiments with pyrene-labeled apoA-I are consistent with the N-terminal helix bundle domain interacting with proteins resident on the HDL particle surface. Dissociation constants (Kd) measured for WT human apoA-I interactions with HDL2 and HDL3 are about 10 µM, indicating that the binding is low affinity. This Kd value does not apply to all of the apoA-I molecules on the HDL particle but only to a relatively small, labile pool. |
| first_indexed | 2024-12-17T23:34:02Z |
| format | Article |
| id | doaj.art-e6e4b58ccd8447ce995551152eb7c5c8 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T23:34:02Z |
| publishDate | 2010-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-e6e4b58ccd8447ce995551152eb7c5c82022-12-21T21:28:35ZengElsevierJournal of Lipid Research0022-22752010-03-01513606617Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particlesSissel Lund-Katz0David Nguyen1Padmaja Dhanasekaran2Momoe Kono3Margaret Nickel4Hiroyuki Saito5Michael C. Phillips6Lipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318Lipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318Lipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318Department of Biophysical Chemistry, Kobe Pharmaceutical University, Kobe 658-8558, JapanLipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318Lipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318To whom correspondence should be addressed; Lipid Research Group, Division of Gastroenterology, Hepatology, and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4318The partitioning of apolipoprotein A-I (apoA-I) molecules in plasma between HDL-bound and -unbound states is an integral part of HDL metabolism. We used the surface plasmon resonance (SPR) technique to monitor in real time the reversible binding of apoA-I to HDL. Biotinylated human HDL2 and HDL3 were immobilized on a streptavidin-coated SPR sensor chip, and apoA-I solutions at different concentrations were flowed across the surface. The wild-type (WT) human and mouse apoA-I/HDL interaction involves a two-step process; apoA-I initially binds to HDL with fast association and dissociation rates, followed by a step exhibiting slower kinetics. The isolated N-terminal helix bundle domains of human and mouse apoA-I also exhibit a two-step binding process, consistent with the second slower step involving opening of the helix bundle domain. The results of fluorescence experiments with pyrene-labeled apoA-I are consistent with the N-terminal helix bundle domain interacting with proteins resident on the HDL particle surface. Dissociation constants (Kd) measured for WT human apoA-I interactions with HDL2 and HDL3 are about 10 µM, indicating that the binding is low affinity. This Kd value does not apply to all of the apoA-I molecules on the HDL particle but only to a relatively small, labile pool.http://www.sciencedirect.com/science/article/pii/S0022227520305186 |
| spellingShingle | Sissel Lund-Katz David Nguyen Padmaja Dhanasekaran Momoe Kono Margaret Nickel Hiroyuki Saito Michael C. Phillips Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles Journal of Lipid Research |
| title | Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles |
| title_full | Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles |
| title_fullStr | Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles |
| title_full_unstemmed | Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles |
| title_short | Surface plasmon resonance analysis of the mechanism of binding of apoA-I to high density lipoprotein particles |
| title_sort | surface plasmon resonance analysis of the mechanism of binding of apoa i to high density lipoprotein particles |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520305186 |
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