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Description

Oxidatively damaged apolipoprotein A1 (apoA1, the main protein constituent of high density lipoprotein (HDL) - the “good cholesterol”) has been isolated from circulating plasma and atherosclerosis plaque with modification of lysine amino acid residues to homocittruline (carbamyllysine) in apoA1 identified as a modification found increased in smokers that correlates with elevated cardiovascular risk for myocardial infarction (MI), stoke or death. The selected carbamyllysine tRNA synthetase will then be used to express a series of human apoA1 alleles containing amber codons in place of various lysine residues and assayed for its ability to form nascent HDL particles and to activate NF-κB and vascular adhesion molecule expression on endothelial cells.

Individual clones from this round of selection will be isolated and tested the clone encoding the synthase which best incorporates carbamyllysine at the lowest uAA concentration. The selected carbamyllysine tRNA synthetase will then be used to express a series of human apoA1 alleles containing amber codons in place of various lysine residues and assayed for its ability to form nascent HDL particles and to activate NF-κB and vascular adhesion molecule expression on endothelial cells.

Publication Date

9-4-2014

Disciplines

Medical Biochemistry

Selection of an Eevolved Orthogonal Ne-Acetyllysine tRNAsynthetase that can Incorporate the Unnantural Amino Acid L-homocitrulline Site-specifically into Proteins

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