title: Phosphorylated human galectin-3: facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy.
abstract: Galectin-3 has a unique modular design. Its short N-terminal stretch can be phosphorylated, relevant for nuclear export and anti-anoikis/apoptosis activity. Enzymatic modification by casein kinase 1 at constant ATP concentration yielded mg quantities of mono- and diphosphorylated derivatives at Ser5/Ser11 in a 2:1 ratio. Their carbohydrate-inhibitable binding to asialofetuin, cell surfaces of three tumor lines, rabbit erythrocytes leading to haemagglutination and cytoplasmic sites in fixed tissue sections was not markedly altered relative to phosphate-free galectin-3. Spectroscopically, phosphorylation induced alterations in the far UV CD, indicative of an increase in ordered structure. This is accompanied by changes in the environment of aromatic amino acids signified by shifts in the near UV CD.
authors.affiliation: Biomolecular Interactions, German Cancer Research Center, Heidelberg, Germany. d.kuebler@dkfz.de
authors.forename: Dieter
authors.initials: D
authors.lastname: Kübler
authors.forename: Chien-Wen
authors.initials: CW
authors.lastname: Hung
authors.forename: Tarun K
authors.initials: TK
authors.lastname: Dam
authors.forename: Jürgen
authors.initials: J
authors.lastname: Kopitz
authors.forename: Sabine
authors.initials: S
authors.lastname: André
authors.forename: Herbert
authors.initials: H
authors.lastname: Kaltner
authors.forename: Michaela
authors.initials: M
authors.lastname: Lohr
authors.forename: Joachim C
authors.initials: JC
authors.lastname: Manning
authors.forename: Lizhong
authors.initials: L
authors.lastname: He
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Anton
authors.initials: A
authors.lastname: Middelberg
authors.forename: C Fred
authors.initials: CF
authors.lastname: Brewer
authors.forename: Jennifer
authors.initials: J
authors.lastname: Reed
authors.forename: Wolf-Dieter
authors.initials: WD
authors.lastname: Lehmann
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Gabius
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001212:Asialoglycoproteins / Q000378:metabolism; D047389:Casein Kinase I / Q000378:metabolism; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D060748:Fetuins; D005434:Flow Cytometry; D037502:Galectin 3 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007583:Jejunum / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D007785:Lactose / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D012694:Serine / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D001212:Asialoglycoproteins; D060748:Fetuins; D037502:Galectin 3; D000509:alpha-Fetoproteins; C017029:asialofetuin; D012694:Serine; D000255:Adenosine Triphosphate; D047389:Casein Kinase I; D007785:Lactose