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Article10.1161/circ.116.suppl_16.ii_300

Abstract 1456: Proteomic Analysis Of Proteins Bound By D-4F In Plasma From C57bl/6 And ApoA-I KO Mice

Hao Xu,Kirkwood A. Pritchard,Basam Wakim,Michael Olivier,Yue‐De Yang-2007-10-16-Circulation
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TL;DRAbstract

D-4F, an 18 amino acid apolipoprotein A-I (ApoA-I) mimetic, has been shown to be highly effective at improving vascular function in murine models of atherosclerosis. It has been hypothesized that D-4F inhibits atherosclerosis and increases vasodilation by interacting with ApoA-I to improve HDL function. However, the mechanisms by which D-4F protects vascular function remain unclear. We hypothesize that D-4F binds proteins involved in atherosclerosis. To test this hypothesis, we incubated HA-tagged D-4F with plasma from C57bl/6 and ApoA-I knockout mice (ApoA-I KO) on a C57bl/6 background and identified the proteins that were bound. Plasma from C57bl/6 mice or ApoA-I KO mice (n=6/group) was incubated with HA-GG-D-4F (Ac-YPYDVPDYA-GG-DWFKAFYDKVAEKFKEAF-NH2) or HA-GG-scramble-D-4F (Ac-YPYDVPDYA-GG-DWFAKDYFKKAFVEEFAK-NH2) at 10μg/ml. Next an anti-HA antibody was used to immunoprecipitate the HA-tagged peptides and associated proteins. The proteins were then analyzed by ESI-nano-LC/MS/MS. An

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D-4F, an 18 amino acid apolipoprotein A-I (ApoA-I) mimetic, has been shown to be highly effective at improving vascular function in murine models of atherosclerosis. It has been hypothesized that D-4F inhibits atherosclerosis and increases vasodilation by interacting with ApoA-I to improve HDL function. However, the mechanisms by which D-4F protects vascular function remain unclear. We hypothesize that D-4F binds proteins involved in atherosclerosis. To test this hypothesis, we incubated HA-tagged D-4F with plasma from C57bl/6 and ApoA-I knockout mice (ApoA-I KO) on a C57bl/6 background and identified the proteins that were bound. Plasma from C57bl/6 mice or ApoA-I KO mice (n=6/group) was incubated with HA-GG-D-4F (Ac-YPYDVPDYA-GG-DWFKAFYDKVAEKFKEAF-NH2) or HA-GG-scramble-D-4F (Ac-YPYDVPDYA-GG-DWFAKDYFKKAFVEEFAK-NH2) at 10μg/ml. Next an anti-HA antibody was used to immunoprecipitate the HA-tagged peptides and associated proteins. The proteins were then analyzed by ESI-nano-LC/MS/MS. An

Keywords

SerineApolipoprotein BApolipoprotein EKnockout mouseBiochemistryProteasesMolecular biologyMedicine

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