title: Trafficking of platelet-activating factor acetylhydrolase type II in response to oxidative stress.
abstract: Platelet-activating factor acetylhydrolase type II (PAFAH-II) is an intracellular phospholipase A(2) enzyme that hydrolyzes platelet-activating factor and oxidatively fragmented phospholipids. This N-terminally myristoylated protein becomes associated with cytoplasm-facing cell membranes under oxidative stress. The structural requirements for binding of PAFAH-II to membranes in response to oxidative stress are unknown. To begin elucidating the mechanism of trafficking and stress response, we constructed a homology model of PAFAH-II. From the predicted membrane orientation of PAFAH-II, the N-terminal myristoyl group and a hydrophobic patch are hypothesized to be involved in membrane binding. Localization studies of human PAFAH-II in HEK293 cells indicated that an unmyristoylated mutant remained cytoplasmic under stressed and unstressed conditions. The myristoylated wild-type enzyme was partially localized to the cytoplasmic membranes prior to stress and became more localized to these membranes upon stress. A triple mutation of three hydrophobic patch residues of the membrane binding region likewise did not localize to membranes following stress. These results indicate that both the myristoyl group and the hydrophobic patch are essential for proper trafficking of the enzyme to the membranes following oxidative stress. Additionally, colocalization studies using organelle-specific proteins demonstrate that PAFAH-II is transported to the membranes of both the endoplasmic reticulum and Golgi apparatus.
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716, United States.
authors: Anastasia F
authors: AF
authors: Thévenin
authors: Elizabeth S
authors: ES
authors: Monillas
authors: Jason M
authors: JM
authors: Winget
authors: Kirk
authors: K
authors: Czymmek
authors: Brian J
authors: BJ
authors: Bahnson
mesh_terms: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000201:enzymology; D006056:Golgi Apparatus / Q000201:enzymology; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D018384:Oxidative Stress / Q000502:physiology*
chemical_list: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase