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Biochim Biophys Acta. 2008 Apr;1780(4):716-22. doi: 10.1016/j.bbagen.2008.01.018. Epub 2008 Feb 12.

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

Author information

1
Biomolecular Interactions, German Cancer Research Center, Heidelberg, Germany. d.kuebler@dkfz.de

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.

PMID:
18302943
DOI:
10.1016/j.bbagen.2008.01.018
[Indexed for MEDLINE]

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