Ghrelin Octanoylation Is Completely Stabilized in Biological Samples by Alkyl Fluorophosphonates
| Autor: | Michelle A. Sieburg, Joseph E. Darling, Trevor Rodrigues, James L. Hougland, Alfonso Abizaid, Kayleigh R. McGovern-Gooch |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine medicine.medical_specialty Acylation 030209 endocrinology & metabolism Peptide hormone Biology Energy homeostasis Phosphates Serine Fluorides 03 medical and health sciences 0302 clinical medicine Endocrinology Blood serum Internal medicine medicine Animals Humans Rats Wistar Receptor chemistry.chemical_classification Protein Stability digestive oral and skin physiology Ghrelin Rats HEK293 Cells 030104 developmental biology Enzyme chemistry Biochemistry Acyltransferases hormones hormone substitutes and hormone antagonists Hormone |
| Zdroj: | Endocrinology. 157:4330-4338 |
| ISSN: | 1945-7170 0013-7227 |
| DOI: | 10.1210/en.2016-1657 |
| Popis: | Ghrelin is a peptide hormone involved in multiple physiological processes related to energy homeostasis. This hormone features a unique posttranslational serine octanoylation modification catalyzed by the enzyme ghrelin O-acyltransferase, with serine octanoylation essential for ghrelin to bind and activate its cognate receptor. Ghrelin deacylation rapidly occurs in circulation, with both ghrelin and desacyl ghrelin playing important roles in biological signaling. Understanding the regulation and physiological impact of ghrelin signaling requires the ability to rapidly protect ghrelin from deacylation in biological samples such as blood serum or cell lysates to preserve the relative concentrations of ghrelin and desacyl ghrelin. In in vitro ghrelin O-acyltransferase activity assays using insect microsomal protein fractions and mammalian cell lysate and blood serum, we demonstrate that alkyl fluorophosphonate treatment provides rapid, complete, and long-lasting protection of ghrelin acylation against serine ester hydrolysis without interference in enzyme assay or ELISA analysis. Our results support alkyl fluorophosphonate treatment as a general tool for stabilizing ghrelin and improving measurement of ghrelin and desacyl ghrelin concentrations in biochemical and clinical investigations and suggest current estimates for active ghrelin concentration and the ghrelin to desacyl ghrelin ratio in circulation may underestimate in vivo conditions. |
| Databáze: | OpenAIRE |
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