Biochemical assays for ghrelin acylation and inhibition of ghrelin O-acyltransferase

Michelle A. Sieburg, Elizabeth R. Cleverdon, James L Hougland

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Ghrelin O-acyltransferase (GOAT) is an enzyme responsible for octanoylating and activating ghrelin, a peptide hormone that plays a key role in energy regulation and hunger signaling. Due to its nature as an integral membrane protein, GOAT has yet to be purified in active form which has complicated biochemical and structural studies of GOAT-catalyzed ghrelin acylation. In this chapter, we describe protocols for efficient expression and enrichment of GOAT in insect cell-derived microsomal fraction, HPLC-based assays for GOAT acylation activity employing fluorescently labeled peptides, and assessment of inhibitor potency against GOAT.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages227-241
Number of pages15
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume2009
ISSN (Print)1064-3745

Keywords

  • Enzyme inhibition
  • Ghrelin
  • Ghrelin O-acyltransferase (GOAT)
  • HPLC
  • Membrane-bound O-acyltransferase (MBOAT)
  • Protein acylation
  • Protein octanoylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Sieburg, M. A., Cleverdon, E. R., & Hougland, J. L. (2019). Biochemical assays for ghrelin acylation and inhibition of ghrelin O-acyltransferase. In Methods in Molecular Biology (pp. 227-241). (Methods in Molecular Biology; Vol. 2009). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9532-5_18