Mining the O-mannose glycoproteome reveals cadherins as major O-mannosylated glycoproteins

TY - JOUR

T1 - Mining the O-mannose glycoproteome reveals cadherins as major O-mannosylated glycoproteins

AU - Vester-Christensen, Malene B

AU - Halim, Adnan

AU - Joshi, Hiren Jitendra

AU - Steentoft, Catharina

AU - Bennett, Eric P

AU - Levery, Steven B

AU - Vakhrushev, Sergey Y

AU - Clausen, Henrik

PY - 2013/12/24

Y1 - 2013/12/24

N2 - The metazoan O-mannose (O-Man) glycoproteome is largely unknown. It has been shown that up to 30% of brain O-glycans are of the O-Man type, but essentially only alpha-dystroglycan (α-DG) of the dystrophin-glycoprotein complex is well characterized as an O-Man glycoprotein. Defects in O-Man glycosylation underlie congenital muscular dystrophies and considerable efforts have been devoted to explore this O-glycoproteome without much success. Here, we used our SimpleCell strategy using nuclease-mediated gene editing of a human cell line (MDA-MB-231) to reduce the structural heterogeneity of O-Man glycans and to probe the O-Man glycoproteome. In this breast cancer cell line we found that O-Man glycosylation is primarily found on cadherins and plexins on β-strands in extracellular cadherin and Ig-like, plexin and transcription factor domains. The positions and evolutionary conservation of O-Man glycans in cadherins suggest that they play important functional roles for this large group of cell adhesion glycoproteins, which can now be addressed. The developed O-Man SimpleCell strategy is applicable to most types of cell lines and enables proteome-wide discovery of O-Man protein glycosylation.

AB - The metazoan O-mannose (O-Man) glycoproteome is largely unknown. It has been shown that up to 30% of brain O-glycans are of the O-Man type, but essentially only alpha-dystroglycan (α-DG) of the dystrophin-glycoprotein complex is well characterized as an O-Man glycoprotein. Defects in O-Man glycosylation underlie congenital muscular dystrophies and considerable efforts have been devoted to explore this O-glycoproteome without much success. Here, we used our SimpleCell strategy using nuclease-mediated gene editing of a human cell line (MDA-MB-231) to reduce the structural heterogeneity of O-Man glycans and to probe the O-Man glycoproteome. In this breast cancer cell line we found that O-Man glycosylation is primarily found on cadherins and plexins on β-strands in extracellular cadherin and Ig-like, plexin and transcription factor domains. The positions and evolutionary conservation of O-Man glycans in cadherins suggest that they play important functional roles for this large group of cell adhesion glycoproteins, which can now be addressed. The developed O-Man SimpleCell strategy is applicable to most types of cell lines and enables proteome-wide discovery of O-Man protein glycosylation.

KW - Cadherins

KW - Cell Adhesion Molecules

KW - Cell Line, Tumor

KW - Glycoproteins

KW - Glycosylation

KW - Humans

KW - Mannose

KW - Mass Spectrometry

KW - Nerve Tissue Proteins

KW - Proteome

KW - Proteomics

U2 - 10.1073/pnas.1313446110

DO - 10.1073/pnas.1313446110

M3 - Journal article

C2 - 24101494

SN - 0027-8424

VL - 110

SP - 21018

EP - 21023

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 52

ER -