Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue

Morris A. Kostiuk, Maria M. Corvi, Bernd O. Keller, Greg Plummer, Jennifer A. Prescher, Matthew J. Hangauer, Carolyn R. Bertozzi, Gurram Rajaiah, John R. Falck, Luc G. Berthiaume

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

Increased levels of circulating saturated free fatty acids, such as palmitate, have been implicated in the etiology of type II diabetes and cancer. In addition to being a constituent of glycerolipids and a source of energy, palmitate also covalently attaches to numerous cellular proteins via a process named palmitoylation. Recognized for its roles in membrane tethering, cellular signaling, and protein trafficking, palmitoylation is also emerging as a potential regulator of metabolism. Indeed, we showed previously that the acylation of two mitochondrial proteins at their active site cysteine residues result in their inhibition. Herein, we sought to identify other palmitoylated proteins in mitochondria using a nonradioactive bio-orthogonal azido-palmitate analog that can be selectively derivatized with various tagged triarylphosphines. Our results show that, like palmitate, incorporation of azido-palmitate occurred on mitochondrial proteins via thioester bonds at sites that could be competed out by palmitoyl-CoA. Using this method, we identified 21 putative palmitoylated proteins in the rat liver mitochondrial matrix, a compartment not recognized for its content in palmitoylated proteins, and confirmed the palmitoylation of newly identified mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase. We postulate that covalent modification and perhaps inhibition of various mitochondrial enzymes by palmitoyl-CoA could lead to the metabolic impairments found in obesity-related diseases.

Original languageEnglish (US)
Pages (from-to)721-732
Number of pages12
JournalFASEB Journal
Volume22
Issue number3
DOIs
StatePublished - Mar 2008

Fingerprint

Palmitates
palmitates
Mitochondrial Proteins
Lipoylation
Palmitoyl Coenzyme A
Proteins
proteins
obesity-related diseases
Cell signaling
Acylation
cell communication
acylation
Mitochondria
protein transport
Protein Transport
Medical problems
Nonesterified Fatty Acids
noninsulin-dependent diabetes mellitus
Metabolism
active sites

Keywords

  • 3-hydroxy-3-methylglutaryl CoA synthase
  • Azido-fatty acid
  • Carbamoyl phosphate synthetase 1
  • Staudinger ligation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Kostiuk, M. A., Corvi, M. M., Keller, B. O., Plummer, G., Prescher, J. A., Hangauer, M. J., ... Berthiaume, L. G. (2008). Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. FASEB Journal, 22(3), 721-732. https://doi.org/10.1096/fj.07-9199com

Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. / Kostiuk, Morris A.; Corvi, Maria M.; Keller, Bernd O.; Plummer, Greg; Prescher, Jennifer A.; Hangauer, Matthew J.; Bertozzi, Carolyn R.; Rajaiah, Gurram; Falck, John R.; Berthiaume, Luc G.

In: FASEB Journal, Vol. 22, No. 3, 03.2008, p. 721-732.

Research output: Contribution to journalArticle

Kostiuk, MA, Corvi, MM, Keller, BO, Plummer, G, Prescher, JA, Hangauer, MJ, Bertozzi, CR, Rajaiah, G, Falck, JR & Berthiaume, LG 2008, 'Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue', FASEB Journal, vol. 22, no. 3, pp. 721-732. https://doi.org/10.1096/fj.07-9199com
Kostiuk MA, Corvi MM, Keller BO, Plummer G, Prescher JA, Hangauer MJ et al. Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. FASEB Journal. 2008 Mar;22(3):721-732. https://doi.org/10.1096/fj.07-9199com
Kostiuk, Morris A. ; Corvi, Maria M. ; Keller, Bernd O. ; Plummer, Greg ; Prescher, Jennifer A. ; Hangauer, Matthew J. ; Bertozzi, Carolyn R. ; Rajaiah, Gurram ; Falck, John R. ; Berthiaume, Luc G. / Identification of palmitoylated mitochondrial proteins using a bio-orthogonal azido-palmitate analogue. In: FASEB Journal. 2008 ; Vol. 22, No. 3. pp. 721-732.
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