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

doi:10.1016/s0021-9258(18)82137-8

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 journal › Article › peer-review

56 Scopus citations

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 language	English (US)
Pages (from-to)	11394-11400
Number of pages	7
Journal	Journal of Biological Chemistry
Volume	268
Issue number	15
DOIs	https://doi.org/10.1016/s0021-9258(18)82137-8
State	Published - May 25 1993
Externally published	Yes

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

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10.1016/s0021-9258(18)82137-8

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@article{451edb5a3f8d4374a6fe21b1aa4e0cc9,

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

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.",

author = "Tetsuro Fujimoto and Eric Stroud and Whatley, \{Ralph E.\} and Prescott, \{Stephen M.\} and Laszlo Muszbek and Michael Laposata and McEver, \{Rodger P.\}",

year = "1993",

month = may,

day = "25",

doi = "10.1016/s0021-9258(18)82137-8",

language = "English (US)",

volume = "268",

pages = "11394--11400",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology Inc.",

number = "15",

}

TY - JOUR

T1 - P-selectin is acylated with palmitic acid and stearic acid at cysteine 766 through a thioester linkage

AU - Fujimoto, Tetsuro

AU - Stroud, Eric

AU - Whatley, Ralph E.

AU - Prescott, Stephen M.

AU - Muszbek, Laszlo

AU - Laposata, Michael

AU - McEver, Rodger P.

PY - 1993/5/25

Y1 - 1993/5/25

N2 - 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.

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.

UR - https://www.scopus.com/pages/publications/0027233704

UR - https://www.scopus.com/inward/citedby.url?scp=0027233704&partnerID=8YFLogxK

U2 - 10.1016/s0021-9258(18)82137-8

DO - 10.1016/s0021-9258(18)82137-8

M3 - Article

C2 - 7684381

AN - SCOPUS:0027233704

SN - 0021-9258

VL - 268

SP - 11394

EP - 11400

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 15

ER -

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

Abstract

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