Title | Endothelial cell palmitoylproteomic identifies novel lipid-modified targets and potential substrates for protein acyl transferases. |
Publication Type | Journal Article |
Year of Publication | 2012 |
Authors | Marin EP, Derakhshan B, Lam TKT, Davalos A, Sessa WC |
Journal | Circ Res |
Volume | 110 |
Issue | 10 |
Pagination | 1336-44 |
Date Published | 2012 May 11 |
ISSN | 1524-4571 |
Keywords | Acetyltransferases, Acyltransferases, Amino Acid Sequence, Animals, Cercopithecus aethiops, COS Cells, Endothelial Cells, HEK293 Cells, Human Umbilical Vein Endothelial Cells, Humans, Lipoylation, Molecular Sequence Data, Platelet Endothelial Cell Adhesion Molecule-1, Proteomics, RNA, Small Interfering, Substrate Specificity, Superoxide Dismutase, Superoxide Dismutase-1 |
Abstract | RATIONALE: Protein S-palmitoylation is the posttranslational attachment of a saturated 16-carbon palmitic acid to a cysteine side chain via a thioester bond. Palmitoylation can affect protein localization, trafficking, stability, and function. The extent and roles of palmitoylation in endothelial cell (EC) biology is not well-understood, partly because of technological limits on palmitoylprotein detection. OBJECTIVE: To develop a method using acyl-biotinyl exchange technology coupled with mass spectrometry to globally isolate and identify palmitoylproteins in ECs. METHODS AND RESULTS: More than 150 putative palmitoyl proteins were identified in ECs using acyl-biotinyl exchange and mass spectrometry. Among the novel palmitoylproteins identified is superoxide dismutase-1, an intensively studied enzyme that protects all cells from oxidative damage. Mutation of cysteine-6 prevents palmitoylation, leads to reduction in superoxide dismutase-1 activity in vivo and in vitro, and inhibits nuclear localization, thereby supporting a functional role for superoxide dismutase-1 palmitoylation. Moreover, we used acyl-biotinyl exchange to search for substrates of particular protein acyl transferases in ECs. We found that palmitoylation of the cell adhesion protein platelet endothelial cell adhesion molecule-1 is dependent on the protein acyl transferase ZDHHC21. We show that knockdown of ZDHHC21 leads to reduced levels of platelet endothelial cell adhesion molecule-1 at the cell surface. CONCLUSIONS: Our data demonstrate the utility of EC palmitoylproteomics to reveal new insights into the role of this important posttranslational lipid modification in EC biology. |
DOI | 10.1161/CIRCRESAHA.112.269514 |
Alternate Journal | Circ. Res. |
PubMed ID | 22496122 |
PubMed Central ID | PMC3428238 |
Grant List | P01 HL070295 / HL / NHLBI NIH HHS / United States HL096670 / HL / NHLBI NIH HHS / United States P01 HL107205 / HL / NHLBI NIH HHS / United States R01 HL61371 / HL / NHLBI NIH HHS / United States R01 HL64793 / HL / NHLBI NIH HHS / United States R01 HL064793 / HL / NHLBI NIH HHS / United States T32 DK007276 / DK / NIDDK NIH HHS / United States N01-HV-28186 / HV / NHLBI NIH HHS / United States K08 HL103831 / HL / NHLBI NIH HHS / United States N01HV28186 / HL / NHLBI NIH HHS / United States 5T32DK007276-33 / DK / NIDDK NIH HHS / United States R37 HL061371 / HL / NHLBI NIH HHS / United States R01 HL096670 / HL / NHLBI NIH HHS / United States P01 HL70295 / HL / NHLBI NIH HHS / United States R01 HL081190 / HL / NHLBI NIH HHS / United States R01 HL061371 / HL / NHLBI NIH HHS / United States |
Submitted by kej2006 on June 6, 2018 - 4:11pm