Notum deacylates Wnt proteins to suppress signalling activity.
Signalling by Wnt proteins is finely balanced to ensure normal development and tissue homeostasis while avoiding diseases such as cancer. This is achieved in part by Notum, a highly conserved secreted feedback antagonist. Notum has been thought to act as a phospholipase, shedding glypicans and associated Wnt proteins from the cell surface. However, this view fails to explain specificity, as glypicans bind many extracellular ligands. Here we provide genetic evidence in Drosophila that Notum requires glypicans to suppress Wnt signalling, but does not cleave their glycophosphatidylinositol anchor. Structural analyses reveal glycosaminoglycan binding sites on Notum, which probably help Notum to co-localize with Wnt proteins. They also identify, at the active site of human and Drosophila Notum, a large hydrophobic pocket that accommodates palmitoleate. Kinetic and mass spectrometric analyses of human proteins show that Notum is a carboxylesterase that removes an essential palmitoleate moiety from Wnt proteins and thus constitutes the first known extracellular protein deacylase.
MRC's National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Satoshi
S
Kakugawa
MRC's National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Paul F
PF
Langton
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
Matthias
M
Zebisch
MRC's National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Steve
S
Howell
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
Tao-Hsin
TH
Chang
Glycosciences Laboratory, Imperial College London, Department of Medicine Du Cane Road, London, W12 0NN UK.
Yan
Y
Liu
Glycosciences Laboratory, Imperial College London, Department of Medicine Du Cane Road, London, W12 0NN UK.
Ten
T
Feizi
Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
Ganka
G
Bineva
Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
Nicola
N
O'Reilly
Cancer Research UK, Clare Hall Laboratories, Blanche Lane, South Mimms, Potters Bar, Hertfordshire. EN6 3LD, UK.
Ambrosius P
AP
Snijders
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
E Yvonne
EY
Jones
MRC's National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Jean-Paul
JP
Vincent
D000215:Acylation; D000818:Animals; D001665:Binding Sites; D043182:Carboxylesterase / Q000737:chemistry / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism*; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005229:Fatty Acids, Monounsaturated / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D053673:Glypicans / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008024:Ligands; D013058:Mass Spectrometry; D008958:Models, Molecular; D011485:Protein Binding; D051153:Wnt Proteins / Q000737:chemistry* / Q000378:metabolism*; D060449:Wnt Signaling Pathway
D029721:Drosophila Proteins; D005229:Fatty Acids, Monounsaturated; D017261:Glycosylphosphatidylinositols; D053673:Glypicans; D008024:Ligands; C456459:Notum protein, Drosophila; D051153:Wnt Proteins; C008757:palmitoleic acid; D004950:Esterases; C000596390:Notum protein, human; D043182:Carboxylesterase