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

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

Fingerprint

Acyltransferases
Ghrelin
Acylation
Hunger
Inhibition (Psychology)
Peptide Hormones
Insects
Membrane Proteins
High Pressure Liquid Chromatography
Peptides
Enzymes

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

Cite this

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

Biochemical assays for ghrelin acylation and inhibition of ghrelin O-acyltransferase. / Sieburg, Michelle A.; Cleverdon, Elizabeth R.; Hougland, James L.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 227-241 (Methods in Molecular Biology; Vol. 2009).

Research output: Chapter in Book/Report/Conference proceedingChapter

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