P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage

Tetsuro Fujimoto, Eric Stroud, Ralph E. Whatley, Stephen M. Prescott, Laszlo Muszbek, Michael Laposata, Rodger P. McEver

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

We report that the adhesion receptor P-selectin can be metabolically labeled with [3H]palmitic acid in human platelets. Analysis of alkaline methanolysis products from labeled protein demonstrated that the radioactivity associated with P-selectin was covalently bound palmitic acid. [3H]Palmitic acid was cleaved by hydroxylamine treatment at neutral pH and by reducing agents, indicating that acylation occurred through a thioester linkage. Both stearic acid and palmitic acid were detected by gas chromatography-mass spectrometry analysis of alkaline hydrolysates of purified P-selectin. Deletion or mutation of Cys766 eliminated [3H] palmitic acid labeling of P-selectin in transfected COS-7 cells. We conclude that the cytoplasmic domain of P-selectin is acylated at Cys766 through a thioester bond. Fatty acid acylation may regulate intracellular trafficking or other functions of P-selectin.

Original languageEnglish (US)
Pages (from-to)11394-11400
Number of pages7
JournalJournal of Biological Chemistry
Volume268
Issue number15
StatePublished - May 25 1993
Externally publishedYes

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P-Selectin
Palmitic Acid
Cysteine
Acylation
Hydroxylamine
Sequence Deletion
COS Cells
Reducing Agents
Radioactivity
Platelets
Gas chromatography
Gas Chromatography-Mass Spectrometry
Labeling
Mass spectrometry
stearic acid
Fatty Acids
Blood Platelets
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fujimoto, T., Stroud, E., Whatley, R. E., Prescott, S. M., Muszbek, L., Laposata, M., & McEver, R. P. (1993). P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage. Journal of Biological Chemistry, 268(15), 11394-11400.

P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage. / Fujimoto, Tetsuro; Stroud, Eric; Whatley, Ralph E.; Prescott, Stephen M.; Muszbek, Laszlo; Laposata, Michael; McEver, Rodger P.

In: Journal of Biological Chemistry, Vol. 268, No. 15, 25.05.1993, p. 11394-11400.

Research output: Contribution to journalArticle

Fujimoto, T, Stroud, E, Whatley, RE, Prescott, SM, Muszbek, L, Laposata, M & McEver, RP 1993, 'P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage', Journal of Biological Chemistry, vol. 268, no. 15, pp. 11394-11400.
Fujimoto T, Stroud E, Whatley RE, Prescott SM, Muszbek L, Laposata M et al. P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage. Journal of Biological Chemistry. 1993 May 25;268(15):11394-11400.
Fujimoto, Tetsuro ; Stroud, Eric ; Whatley, Ralph E. ; Prescott, Stephen M. ; Muszbek, Laszlo ; Laposata, Michael ; McEver, Rodger P. / P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 15. pp. 11394-11400.
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AB - We report that the adhesion receptor P-selectin can be metabolically labeled with [3H]palmitic acid in human platelets. Analysis of alkaline methanolysis products from labeled protein demonstrated that the radioactivity associated with P-selectin was covalently bound palmitic acid. [3H]Palmitic acid was cleaved by hydroxylamine treatment at neutral pH and by reducing agents, indicating that acylation occurred through a thioester linkage. Both stearic acid and palmitic acid were detected by gas chromatography-mass spectrometry analysis of alkaline hydrolysates of purified P-selectin. Deletion or mutation of Cys766 eliminated [3H] palmitic acid labeling of P-selectin in transfected COS-7 cells. We conclude that the cytoplasmic domain of P-selectin is acylated at Cys766 through a thioester bond. Fatty acid acylation may regulate intracellular trafficking or other functions of P-selectin.

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