title: Studying N-linked glycosylation of receptor tyrosine kinases.
abstract: Metabolic alterations have been identified as a frequent event in cancer. This is often associated with increased flux through glycolysis, and also a secondary pathway to glycolysis, hexosamine biosynthetic pathway (HBP). HBP provides substrate for N-linked glycosylation, which occurs in the endoplasmic reticulum and the Golgi apparatus. N-linked glycosylation supports protein folding and correct sorting of proteins to plasma membrane and secretion. This process generates complex glycoforms, which can be recognized by other proteins and glycosylation of receptor tyrosine kinases (RTK) can also regulate their plasma-membrane retention time. Of special interest for experimental biologists, plants produce proteins, termed lectins, which bind with high specificity to glyco-conjugates. For the purposes of molecular biology, plant lectins can be conjugated to different moieties, such as agarose beads, which enable precipitation of specifically glycosylated proteins. In this chapter, we describe in detail how to perform pull-down experiments with commercially available lectins to identify changes in the glycosylation of RTKs.
authors.affiliation: Prostate Cancer Research Group, Centre for Molecular Medicine Norway, Nordic EMBL Partnership, University of Oslo and Oslo University Hospital, Forskningsparken, Gaustadalléen 21, Oslo, 0349, Norway, harri.itkonen@ncmm.uio.no.
authors.forename: Harri M
authors.initials: HM
authors.lastname: Itkonen
authors.forename: Ian G
authors.initials: IG
authors.lastname: Mills
mesh_terms: D000728:Androgens / Q000494:pharmacology; D001665:Binding Sites; D015153:Blotting, Western / Q000379:methods*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D006031:Glycosylation; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D010835:Phytohemagglutinins / Q000737:chemistry*; D011467:Prostate / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D017526:Receptor, IGF Type 1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D000728:Androgens; D010835:Phytohemagglutinins; C100156:leukoagglutinins, plants; D017526:Receptor, IGF Type 1