Exocytosis, the fusion of a vesicle to a cellular membrane, involves a protein named SNAP-25. This protein, containing two alpha helices connected with a linker region, is localized to the cell membrane via palmitic acids attached to the cysteine residues of its linker region in a process called palmitoylation. Are cysteine residues of the SNAP-25 linker region palmitoylated in an ordered manner and to a particular extent? The answer to this question may give insight into the regulated nature of exocytosis. While it is generally accepted that SNAP-25 must be palmitoylated in order to perform its exocytotic functions, the details surrounding this process are still being discovered, defined, and understood. In these studies we replicate the oxidation, reduction, and palmitoylation of SNAP-25 in vitro. Palmitoylating SNAP-25 in vitro, a process which occurs regularly in vivo, allows us to determine the extent of palmitoylation. In vitro palmitoylation of SNAP-25 was studied both with and without a native palmitoyl acyl transferase (PAT), DHHC-17, the enzyme to attach palmitic acids to cysteines in the linker region of SNAP-25. These studies were done under a variety of conditions designed to identify (1) components necessary for optimal palmitoylation and (2) extent of palmitoylation with components that mimic native conditions. Palmitoylation is a common modification for a variety of proteins, both soluble and membrane-bound. Like phosphorylation, palmitoylation is reversible and may play an important role in regulation of cellular processes. Specifically, the palmitoylation of SNAP-25 may play a critical role in the regulation of exocytosis and therefore learning further details about this important process may help us to better understand a variety of neurodegenerative diseases and states of decreased or compromised exocytosis.
College and Department
Life Sciences; Physiology and Developmental Biology
BYU ScholarsArchive Citation
DaBell, Alex McGregor, "Extent of Cysteine Modification of SNAP-25 In vitro" (2014). All Theses and Dissertations. 4361.
palmitoylation, palmitoyl acyl transferase, SNAP-25, DHHC-17