Phosphorylated human galectin-3: Facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy

Dieter Kübler, Chien Wen Hung, Tarun K. Dam, Jürgen Kopitz, Sabine André, Herbert Kaltner, Michaela Lohr, Joachim C. Manning, Lizhong He, Hui Wang, Anton Middelberg, C. Fred Brewer, Jennifer Reed, Wolf Dieter Lehmann, Hans Joachim Gabius

Research output: Contribution to journalArticle

36 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)716-722
Number of pages7
JournalBiochimica et Biophysica Acta - General Subjects
Volume1780
Issue number4
DOIs
StatePublished - Apr 1 2008

Keywords

  • Apoptosis
  • Histochemistry
  • Lectin
  • Serine phosphorylation
  • Titania chromatography

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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    Kübler, D., Hung, C. W., Dam, T. K., Kopitz, J., André, S., Kaltner, H., Lohr, M., Manning, J. C., He, L., Wang, H., Middelberg, A., Brewer, C. F., Reed, J., Lehmann, W. D., & Gabius, H. J. (2008). Phosphorylated human galectin-3: Facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy. Biochimica et Biophysica Acta - General Subjects, 1780(4), 716-722. https://doi.org/10.1016/j.bbagen.2008.01.018