PUBLICATION
Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling
- Authors
- Liao, W.J., Tsao, K.C., Yang, R.B.
- ID
- ZDB-PUB-151225-9
- Date
- 2016
- Source
- The Biochemical journal 473(5): 661-72 (Journal)
- Registered Authors
- Yang, Ruey-Bing (Ray)
- Keywords
- bone morphogenetic protein, electrostatic interaction, N-glycosylation, peripheral membrane protein, SCUBE1, signalling
- MeSH Terms
-
- Amino Acid Motifs
- Amino Acid Sequence
- Animals
- Bone Morphogenetic Proteins/metabolism*
- Cell Membrane/metabolism
- Glycosylation
- HEK293 Cells
- Hematopoiesis
- Humans
- Membrane Microdomains/metabolism
- Membrane Proteins/genetics
- Membrane Proteins/metabolism*
- Molecular Sequence Data
- Mutation
- Oligosaccharides/metabolism*
- Protein Binding
- Protein Structure, Tertiary
- Signal Transduction
- Static Electricity
- Zebrafish/blood
- PubMed
- 26699903 Full text @ Biochem. J.
Citation
Liao, W.J., Tsao, K.C., Yang, R.B. (2016) Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. The Biochemical journal. 473(5):661-72.
Abstract
SCUBE1, a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an NH2-terminal signal peptide sequence followed by 9 epidermal growth factor (EGF)-like repeats, a spacer region and 3 cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the COOH terminus. Soluble SCUBE1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. However, the mechanism responsible for the membrane tethering and biological importance of N-glycosylation of SCUBE1 remains largely unknown. In this study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. Furthermore, deglycosylation by peptide-N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of hematopoietic and erythroid markers ( scl and gata1 ) in scube1 -knockdown embryos. We describe novel mechanisms in targeting SCUBE1 to the plasma membrane and demonstrate that N-glycans are required for SCUBE1 functions during primitive hematopoiesis in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping