Swift residue-screening identifies key N-glycosylated asparagines sufficient for surface expression of neuroglycoprotein Lingo-1

• Published in: FEBS letters Vol. 583; no. 6; pp. 1034 - 1038
• Main Authors: Zhong, Xiaotian, Pocas, Jennifer, Liu, Yan, Wu, Paul W., Mosyak, Lidia, Somers, Will, Kriz, Ron
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 18.03.2009
• Subjects: a secreted version of human Lingo-1; Antigens, Surface - metabolism; Asparagine - analysis; Asparagine - metabolism; Cell Membrane - metabolism; Cells, Cultured; CMV; cytomegalovirus; Glycoproteins - chemistry; Glycoproteins - metabolism; Glycosylation; Humans; leucine-rich-repeat; LRR; Mammalian transient expression; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Models, Biological; Mutagenesis, Site-Directed - methods; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neuroglycoprotein; PCR; PEI; Plasmids - chemistry; polyethylenimine; polymerase chain reaction; Polymerase Chain Reaction - methods; Protein Conformation; Residue-screening; Sequence Analysis, Protein - methods; sLingo-1; Surface expression; Time Factors; Residue-screening; PEI; Mammalian transient expression; sLingo-1; Surface expression; Glycosylation; Neuroglycoprotein; CMV; LRR; PCR
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2009.02.034

Advances in genomics and proteomics have generated the needs for the efficient identification of key residues for structure and function of target proteins. Here we report the utilization of a new residue-screening approach, which combines a mammalian high-throughput transient expression system with a PCR-based expression cassette, for the study of the post-translational modification. Applying this approach results in a quick identification of essential N-glycosylation sites of a heavily glycosylated neuroglycoprotein Lingo-1, which are sufficient for the support of its surface expression. These key N-glycosylated sites uniquely locate on the concave surface of the elongated arc-shape structure of the leucine-rich repeat domain. The swift residue-screening approach may provide a new strategy for structural and functional analysis.