Multifaceted approaches including neoglycolipid oligosaccharide microarrays to ligand discovery for malectin.
In this chapter, we describe the key procedures for isolation of the oligosaccharides and the preparation of neoglycolipid probes together with expression of malectin that have enabled the discovery of the highly selective binding of this newly described protein in the endoplasmic reticulum (ER) to a diglucosyl high-mannose N-glycan. This is the first indication of a bioactivity for a diglucosyl high-mannose N-glycan of the type that occurs in the ER of eukaryotic cells and which is an intermediate in the early steps of the N-glycosylation pathway of nascent proteins. The malectin story is an example of a powerful convergence of disciplines in biological sciences: (i) developmental biology, (ii) bioinformatics, (iii) recombinant protein expression, (iv) protein structural studies, (v) glucan biochemistry, and (vi) drug-assisted engineering of oligosaccharide biosynthesis, culminating in (vii) oligosaccharide "designer" microarrays, to clinch the remarkable selectivity of the binding of this newly discovered ER protein. Thus, the way is open to the identification of the role of malectin in the N-glycosylation pathway.
Palma
Angelina S
AS
Glycosciences Laboratory, Faculty of Medicine, Imperial College London, Northwick Park Hospital Campus, Harrow, Middlesex, United Kingdom.
Liu
Yan
Y
Muhle-Goll
Claudia
C
Butters
Terry D
TD
Zhang
Yibing
Y
Childs
Robert
R
Chai
Wengang
W
Feizi
Ten
T
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002645:Chickens; D006017:Glycolipids / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008024:Ligands; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment; D015398:Signal Transduction
D006017:Glycolipids; D037102:Lectins; D008024:Ligands; C554484:MLEC protein, human; D008565:Membrane Proteins; D009844:Oligosaccharides; D011994:Recombinant Proteins