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Product no.49-66-62408 mm49-66-626949-66-62419 mm49-66-627049-66-624210 mm49-66-627149-66-624311 mm49-66-627249-66-624412 mm49-66-627349-66-624513 mm49-66-627449-66-624614 mm49-66-627549-66-624715 mm49-66-627649-66-624816mm49-66-627749-66-624917 mm49-66-627849-66-625018 mm49-66-627949-66-625119 mm49-66-628049-66-625220 mm49-66-628149-66-625321 mm49-66-628249-66-625422 mm49-66-628349-66-625523mm49-66-628449-66-625624 mm49-66-628549-66-625725 mm49-66-628649-66-625826 mm49-66-628749-66-625927 mm49-66-628849-66-626028 mm49-66-628949-66-626129 mm49-66-629049-66-626230 mm49-66-629149-66-626331 mm49-66-629249-66-626432 mm49-66-629349-66-626533 mm49-66-629449-66-626634 mm49-66-629549-66-626735 mm49-66-629649-66-626836 mm49-66-6297Additional informationWeight1 lbsBrand NameMilwaukee Tools
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Apolipoprotein (apo) mimetic peptides replicate some aspects of HDL function. We have previously reported the effects of compound ATI-5261 on its ability to replicate many functions of native apo A-I in the process of HDL biogenesis. ATI-5261 induced muscle toxicity in wild type C57Bl/6 mice, increased CPK, ALT and AST and increase in triglyceride (Tg) levels. Aromatic phenylalanine residues on the non-polar face of ATI-5261, together with positively charged arginine residues at the lipid-water interface were responsible for these effects. This information was used to create a novel analog (CS-6253) that was non-toxic. We evaluated this peptide designed from the carboxyl terminus of apo E, in its ability to mimic apo A-I functionality. Our data shows that the lipidated particles generated by incubating cells overexpressing ABCA1 with lipid free CS-6253 enhances the rate of ABCA1 lipid efflux with high affinity interactions with native ABCA1 oligomeric forms and plasma membrane micro-domains. Interaction between ABCA1 and lipid free CS-6253 resulted in formation of nascent HDL-CS-6253 particles that are actively remodeled in plasma. Mature HDL-CS-6253 particles deliver cholesterol to liver cells via SR-BI in-vitro. CS-6253 significantly increases cholesterol efflux in murine macrophages and in human THP-1 macrophage-derived foam cells expressing ABCA1. Addition of CS-6253 to plasma dose-dependently displaced apo A-I from -HDL particles and led to de novo formation of pre-1 HDL that stimulates ABCA1 dependent cholesterol efflux efficiently. When incubated with human plasma CS-6253 was also found to bind with HDL and LDL and promoted the transfer of cholesterol from HDL to LDL predominantly. Our data shows that CS-6253 mimics apo A-I in its ability to promote ABCA1-mediated formation of nascent HDL particles, and enhances formation of pre-1 HDL with increase in the cycling of apo A-I between the pre and -HDL particles in-vitro. These mechanisms are potentially anti-atherogenic.
We carried out experiments on CS-6253 using similar protocols initially, as those reported for ATI-5261. For comparisons of ATI-5261 and CS-6253, vs. controls, experiments were run simultaneously under identical conditions so that specific kinetic parameters could be compared. Methods and results can be found on the on-line Supplementary appendix of this article, S1 Appendix. Where protocols differ, these are presented here.
The CS-6253 peptide was synthesized by (Biosynthesis Inc., Lewisville TX) from all L-amino acids and capped with N-terminal acetyl and carboxyl-terminal amide groups. They form a class A amphipathic -helix common to those found in apo A-I and apo E. CS-6253 is a modification of peptide ATI-5261 with a substitution of phenylalanine for leucine residues, and arginine for citrulline residues. For experiments, lyophilized peptide was dissolved in 10 mM phosphate buffer (pH 7.4) saline (PBS) (150 mM NaCl), filter sterilized (0.2 m Nalgene capsules), and stored at 4C until use.
The mice toxicity study protocol was approved and performed according to Veterans Administration Palo Alto Health Care System Institutional Animal Care and Use Committee (IACUC), approval study number AZH1155. The mice toxicity studies were conducted in chow fed mice (male, C57BL/6) eight to ten-week old with intraperitoneal injection of 300 mg/kg CS-6253 and ATI-5261 peptides and PBS. Peptide CS-6253 was assessed for muscle toxicity with, ATI-5261 as positive and PBS as negative control respectively. Blood was collected using retro-orbital eye bleeding at 4 and 6 hours while the animals were under isoflurane. be457b7860
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