Protein palmitoylation is a reversible post-translational lipid modification that allows regulated tethering of proteins to membranes and many of key proteins of cellular signaling, cancer, neuronal transmission, and membrane trafficking are palmitoylated. Despite its well-recognized importance, palmitoylation remains surprisingly understudied. Palmitoylation is difficult to detect and predict. Furthermore, until quite recently, the enzymology of palmitoyl addition had remained a total black box. The first two palmitoylation enzymes, i.e. protein acyl transferases (PATs), were identified only four years ago by work in yeast. The two identified yeast PATs together pointed towards the DHHC protein family as a possible PAT family. Yeast has seven of these DHHC proteins, while 23 are identifiable from the human genome. To facilitate global approaches to palmitoylation, we have developed a new proteomic methodology that purifies and identifies palmitoyl- proteins from complex protein extracts. This approach has first been used in yeast to identify many new palmitoyl-proteins and also to match the various palmitoyl-proteins with their cognate modifying PAT. A new project will apply the same proteomic technologies towards an understanding of palmitoylation in the mammalian brain. Palmitoylation plays a prominent role both in the regulation of synaptic signaling as well as in neuronal development and plasticity. Like the yeast analyses, these first mammalian analyses likely will highlight unanticipated new roles for palmitoylation in the brain, providing stimulus to future neuroscientific research. In addition, this technology also will be used to identify the neuronal palmitoylation substrates of HIP14, a DHHC PAT that has been implicated as participating in Huntington's disease. A final project will build on a recently discovered role for palmitoylation in the quality control mechanisms that act at the endoplasmic reticulum (ER). Of particular interest will be an investigation of the role of palmitoylation in the quality control mechanisms that underlie pathogenesis of demyelinating, neurodegenerative Pelizaeus- Merzbacher disease. Thus, in addition to expanding basic understanding of the cell biology of this lipid modification, we also hope to gain insight into the role of palmitoylation in neurodegenerative disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM065525-08
Application #
7770874
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Chin, Jean
Project Start
2002-06-01
Project End
2012-02-28
Budget Start
2010-03-01
Budget End
2012-02-28
Support Year
8
Fiscal Year
2010
Total Cost
$280,889
Indirect Cost
Name
Wayne State University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Wan, Junmei; Savas, Jeffrey N; Roth, Amy F et al. (2013) Tracking brain palmitoylation change: predominance of glial change in a mouse model of Huntington's disease. Chem Biol 20:1421-34
Roth, Amy F; Papanayotou, Irene; Davis, Nicholas G (2011) The yeast kinase Yck2 has a tripartite palmitoylation signal. Mol Biol Cell 22:2702-15
Singaraja, Roshni R; Huang, Kun; Sanders, Shaun S et al. (2011) Altered palmitoylation and neuropathological deficits in mice lacking HIP14. Hum Mol Genet 20:3899-909
Huang, Kun; Sanders, Shaun S; Kang, Rujun et al. (2011) Wild-type HTT modulates the enzymatic activity of the neuronal palmitoyl transferase HIP14. Hum Mol Genet 20:3356-65
Papanayotou, Irene; Sun, Beimeng; Roth, Amy F et al. (2010) Protein aggregation induced during glass bead lysis of yeast. Yeast 27:801-16
Conibear, Elizabeth; Davis, Nicholas G (2010) Palmitoylation and depalmitoylation dynamics at a glance. J Cell Sci 123:4007-10
Huang, Kun; Kang, Martin H; Askew, Caitlin et al. (2010) Palmitoylation and function of glial glutamate transporter-1 is reduced in the YAC128 mouse model of Huntington disease. Neurobiol Dis 40:207-15
Kang, Rujun; Wan, Junmei; Arstikaitis, Pamela et al. (2008) Neural palmitoyl-proteomics reveals dynamic synaptic palmitoylation. Nature 456:904-9
Wan, Junmei; Roth, Amy F; Bailey, Aaron O et al. (2007) Palmitoylated proteins: purification and identification. Nat Protoc 2:1573-84
Roth, Amy F; Wan, Junmei; Bailey, Aaron O et al. (2006) Global analysis of protein palmitoylation in yeast. Cell 125:1003-13

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