title: Radiometric assay of ghrelin hydrolase activity and 3H-ghrelin distribution into mouse tissues.
abstract: A high-throughput radiometric assay was developed to characterize enzymatic hydrolysis of ghrelin and to track the peptide's fate in vivo. The assay is based on solvent partitioning of [(3)H]-octanoic acid liberated from [(3)H]-octanoyl ghrelin during enzymatic hydrolysis. This simple and cost-effective method facilitates kinetic analysis of ghrelin hydrolase activity of native and mutated butyrylcholinesterases or carboxylesterases from multiple species. In addition, the assay's high sensitivity facilitates ready evaluation of ghrelin's pharmacokinetics and tissue distribution in mice after i.v. bolus administration of radiolabeled peptide.
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA; Kogod Center on Aging, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: Chen.vicky@mayo.edu.
authors.forename: Vicky Ping
authors.initials: VP
authors.lastname: Chen
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA; Kogod Center on Aging, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: Gao.yang@mayo.edu.
authors.forename: Yang
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA; Kogod Center on Aging, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: Geng.liyi@mayo.edu.
authors.forename: Liyi
authors.initials: L
authors.lastname: Geng
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA; Kogod Center on Aging, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: Brimijoin@mayo.edu.
authors.forename: Stephen
authors.initials: S
authors.lastname: Brimijoin
mesh_terms: D000818:Animals; D004789:Enzyme Activation / Q000502:physiology; D054439:Ghrelin / Q000378:metabolism*; D006801:Humans; D006867:Hydrolases / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011874:Radiometry / Q000379:methods; D014018:Tissue Distribution / Q000502:physiology; D014316:Tritium / Q000378:metabolism*
chemical_list: D054439:Ghrelin; D014316:Tritium; D006867:Hydrolases
keywords: Butyrylcholinesterase; Di-isopropyl fluorophosphates (PubChem CID: 5936); Enzyme kinetics; Ghrelin; Human desacyl-ghrelin (PubChem CID: 71728433); Human ghrelin (PubChem CID: 91668172); Pharmacokinetics; Radiometric assay; Toluene (PubChem CID: 1140); iso-OMPA (PubChem CID: 5420)