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.
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X.H. acknowledges support by NIH (RO1-GM057603 and RO1-AR060359) and by Boston Children’s Hospital Intellectual and Developmental Disabilities Research Center (P30 HD-18655). X.H. is an American Cancer Society Research Professor.