title: Methods for Studying Wnt Protein Modifications/Inactivations by Extracellular Enzymes, Tiki and Notum.
abstract: Wnt proteins are modified and inactivated by two extracellular enzymatic antagonists, Tiki and Notum. Tiki proteins act as membrane-tethered metalloproteases to cleave a fragment from the amino terminus of Wnt proteins. Notum is a Wnt deacylase that removes the lipid modification that is essential for Wnt activities. Here, we provide detailed procedures for preparing enzymatic active Tiki and Notum proteins and the in vitro enzymatic reactions. We also describe a metabolic labeling and click chemistry method for detection of Wnt protein acylation.
authors: The F. M. Kirby Neurobiology Center, Boston Children's Hospital, Department of Neurology, Harvard Medical School, CLS 12064, 3 Blackfan Circle, Boston, MA, 02115, USA. Xinjun.zhang@childrens.harvard.edu.
authors: Xinjun
authors: X
authors: Zhang
authors: The F. M. Kirby Neurobiology Center, Boston Children's Hospital, Department of Neurology, Harvard Medical School, CLS 12064, 3 Blackfan Circle, Boston, MA, 02115, USA. xi.he@childrens.harvard.edu.
authors: Xi
authors: X
authors: He
mesh_terms: D000215:Acylation; D057930:Click Chemistry / Q000379:methods*; D004950:Esterases / Q000737:chemistry; D045726:Metalloproteases / Q000737:chemistry; D008967:Molecular Biology / Q000379:methods*; D011499:Protein Processing, Post-Translational / Q000235:genetics; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D060449:Wnt Signaling Pathway
chemical_list: D051153:Wnt Proteins; D004950:Esterases; D045726:Metalloproteases
keywords: Acylation; Click-chemistry; Deacylase; Metalloprotease; Notum; Tiki; Wnt