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, Curtis F. Brewer, Jennifer Reed, Wolf Dieter Lehmann, Hans Joachim Gabius

Research output: Contribution to journalArticle

35 Citations (Scopus)

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 2008

Fingerprint

Galectin 3
Lectins
Spectrum Analysis
Casein Kinase I
Anoikis
Spectroscopy
Aromatic Amino Acids
Phosphorylation
Cell Nucleus Active Transport
Hemagglutination
Tumors
Adenosine Triphosphate
Erythrocytes
Phosphates
Carbohydrates
Tissue
Apoptosis
Rabbits
Derivatives
Neoplasms

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Phosphorylated human galectin-3 : Facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy. / Kübler, Dieter; Hung, Chien Wen; Dam, Tarun K.; Kopitz, Jürgen; André, Sabine; Kaltner, Herbert; Lohr, Michaela; Manning, Joachim C.; He, Lizhong; Wang, Hui; Middelberg, Anton; Brewer, Curtis F.; Reed, Jennifer; Lehmann, Wolf Dieter; Gabius, Hans Joachim.

In: Biochimica et Biophysica Acta - General Subjects, Vol. 1780, No. 4, 04.2008, p. 716-722.

Research output: Contribution to journalArticle

Kübler, D, Hung, CW, Dam, TK, Kopitz, J, André, S, Kaltner, H, Lohr, M, Manning, JC, He, L, Wang, H, Middelberg, A, Brewer, CF, Reed, J, Lehmann, WD & Gabius, HJ 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, vol. 1780, no. 4, pp. 716-722. https://doi.org/10.1016/j.bbagen.2008.01.018
Kübler, Dieter ; Hung, Chien Wen ; Dam, Tarun K. ; Kopitz, Jürgen ; André, Sabine ; Kaltner, Herbert ; Lohr, Michaela ; Manning, Joachim C. ; He, Lizhong ; Wang, Hui ; Middelberg, Anton ; Brewer, Curtis F. ; Reed, Jennifer ; Lehmann, Wolf Dieter ; Gabius, Hans Joachim. / Phosphorylated human galectin-3 : Facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy. In: Biochimica et Biophysica Acta - General Subjects. 2008 ; Vol. 1780, No. 4. pp. 716-722.
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