Probing lipid-protein adduction with alkynyl surrogates: Application to Smith-Lemli-Opitz syndrome

Katherine Windsor, Thiago C. Genaro-Mattos, Hye Young H. Kim, Wei Liu, Keri A. Tallman, Sayuri Miyamoto, Zeljka Korade, Ned A. Porter

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Lipid modifications aid in regulating (and misregulating) protein function and localization. However, efficient methods to screen for a lipid's ability to modify proteins are not readily available. We present a strategy to identify protein-reactive lipids and apply it to a neurodevelopmental disorder, Smith-Lemli-Opitz syndrome (SLOS). Alkynyl surrogates were synthesized for polyunsaturated fatty acids, phospholipids, cholesterol, 7-dehydrocholesterol (7-DHC), and a 7-DHC-derived oxysterol. To probe for protein- reactive lipids, we used click chemistry to biotinylate the alkynyl tag and detected the lipid-adducted proteins with streptavidin Western blotting. In Neuro2a cells, the trend in amount of protein adduction followed known rates of lipid peroxidation (7-DHC >> arachidonic acid > linoleic acid >> cholesterol), with alkynyl-7-DHC producing the most adduction among alkynyl lipids. 7-DHC reductase-deficient cells, which cannot properly metabolize 7-DHC, exhibited signifi- cantly more alkynyl-7-DHC-protein adduction than control cells. Model studies demonstrated that a 7-DHC peroxidation product covalently modifies proteins. We hypothesize that 7-DHC generates electrophiles that can modify the proteome, contributing to SLOS's complex pathology. These probes and methods would allow for analysis of lipid-modified proteomes in SLOS and other disorders exhibiting 7-DHC accumulation. More broadly, the alkynyl lipid library would facilitate exploration of lipid peroxidation's role in specific biological processes in numerous diseases.

Original languageEnglish (US)
Pages (from-to)2842-2850
Number of pages9
JournalJournal of Lipid Research
Volume54
Issue number10
DOIs
StatePublished - Oct 1 2013

Fingerprint

Smith-Lemli-Opitz Syndrome
Lipids
Proteins
Proteome
Lipid Peroxidation
Cholesterol
Click Chemistry
Biological Phenomena
7-dehydrocholesterol
Aptitude
Streptavidin
Linoleic Acid
Unsaturated Fatty Acids
Arachidonic Acid
Pathology
Phospholipids
Western Blotting

Keywords

  • 7-dehydrocholesterol
  • Cholesterol
  • Lipid electrophiles
  • Lipid peroxidation
  • Oxysterol

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Probing lipid-protein adduction with alkynyl surrogates : Application to Smith-Lemli-Opitz syndrome. / Windsor, Katherine; Genaro-Mattos, Thiago C.; Kim, Hye Young H.; Liu, Wei; Tallman, Keri A.; Miyamoto, Sayuri; Korade, Zeljka; Porter, Ned A.

In: Journal of Lipid Research, Vol. 54, No. 10, 01.10.2013, p. 2842-2850.

Research output: Contribution to journalArticle

Windsor, Katherine ; Genaro-Mattos, Thiago C. ; Kim, Hye Young H. ; Liu, Wei ; Tallman, Keri A. ; Miyamoto, Sayuri ; Korade, Zeljka ; Porter, Ned A. / Probing lipid-protein adduction with alkynyl surrogates : Application to Smith-Lemli-Opitz syndrome. In: Journal of Lipid Research. 2013 ; Vol. 54, No. 10. pp. 2842-2850.
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AU - Liu, Wei

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