Investigation of the mechanism of transfer of alpha-tocopherol by the human alpha-tocopherol transfer protein (h-alpha-TTP).

摘要

The human alpha-tocopherol transfer protein (h-alpha-TTP) is understood to be the entity responsible for the specific retention of alpha-tocopherol (alpha-toc) in human tissues over all other forms of vitamin E obtained from the diet. alpha-Tocopherol is the most biologically active form of vitamin E, and to date has been studied extensively with regard to its antioxidant properties and its role of terminating membrane lipid peroxidation chain reactions. However, information surrounding the distribution of alpha-tocopherol, specifically its delivery to intracellular membranes by alpha-TTP, is still unclear and the molecular factors influencing transfer remain elusive. To investigate the mechanism of ligand transfer by the h-alpha-TTP, a fluorescent analogue of alpha-toc has been used in the development of a fluorescence resonance energy transfer (FRET) assay.; (R)-2,5,7,8-tetramethy1-2-[9-(7-nitro-benzo[1,2,5]oxdiazol-4-ylamino)-nonyl]-chroman-6-ol (NBD-toc) has allowed for the development of the FRET-based ligand transfer assay. This ligand has been utilized in a series of experiments where changes were made to acceptor lipid membrane concentration and composition, as well as to the ionic strength and viscosity of the buffer medium. Such changes have yielded evidence supporting a collisional mechanism of ligand transfer by alpha-TTP, and have brought to light a new line of inquiry pertaining to the nature of the forces governing the collisional transfer interaction.; Through elucidation of the transfer mechanism type, a deeper understanding of the transfer event and the in vivo fate of alpha-tocopherol have been obtained. Furthermore, the results presented here allow for a deeper investigation of the forces controlling the collisional protein-membrane interaction and their effect on the transfer of alpha-toc to membranes. Future investigation in this direction will raise the possibility of a complete understanding of the molecular events surrounding the distribution of alpha-toc within the cell and to the body's tissues.