10.1021/ja502109n.s003
Tao Peng
Tao
Peng
Howard C. Hang
Howard C.
Hang
Bifunctional
Fatty Acid Chemical Reporter for Analyzing
S‑Palmitoylated Membrane Protein–Protein Interactions
in Mammalian Cells
American Chemical Society
2015
membrane proteins
IFITM 3
New strategies
protein complexes
acid chemical reporter
IFITM 3 oligomerization
Bifunctional Fatty
Chemical Reporter
Mammalian CellsStudying
2015-01-21 00:00:00
Dataset
https://acs.figshare.com/articles/dataset/Bifunctional_Fatty_Acid_Chemical_Reporter_for_Analyzing_S_Palmitoylated_Membrane_Protein_Protein_Interactions_in_Mammalian_Cells/2213836
Studying
the functions of S-palmitoylated proteins in cells can
be challenging due to the membrane targeting property and dynamic
nature of protein S-palmitoylation. New strategies are therefore needed
to specifically capture S-palmitoylated protein complexes in cellular
membranes for dissecting their functions <i>in vivo</i>.
Here we present a bifunctional fatty acid chemical reporter, x-alk-16,
which contains an alkyne and a diazirine, for metabolic labeling of
S-palmitoylated proteins and photo-cross-linking of their involved
protein complexes in mammalian cells. We demonstrate that x-alk-16
can be metabolically incorporated into known S-palmitoylated proteins
such as H-Ras and IFITM3, a potent antiviral protein, and induce covalent
cross-linking of IFITM3 oligomerization as well as its specific interactions
with other membrane proteins upon in-cell photoactivation. Moreover,
integration of x-alk-16-induced photo-cross-linking with label-free
quantitative proteomics allows identification of new IFITM3 interacting
proteins.