TY - JOUR
T1 - Phosphorylated human galectin-3
T2 - Facile large-scale preparation of active lectin and detection of structural changes by CD spectroscopy
AU - Kübler, Dieter
AU - Hung, Chien Wen
AU - Dam, Tarun K.
AU - Kopitz, Jürgen
AU - André, Sabine
AU - Kaltner, Herbert
AU - Lohr, Michaela
AU - Manning, Joachim C.
AU - He, Lizhong
AU - Wang, Hui
AU - Middelberg, Anton
AU - Brewer, C. Fred
AU - Reed, Jennifer
AU - Lehmann, Wolf Dieter
AU - Gabius, Hans Joachim
N1 - Funding Information:
We are grateful to Drs. B. Friday and A. Raz for reading the manuscript. This study was generously supported by grants of the initiative LMUexcellent and EC Marie Curie Research Training Network (2005-019561).
PY - 2008/4
Y1 - 2008/4
N2 - 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.
AB - 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.
KW - Apoptosis
KW - Histochemistry
KW - Lectin
KW - Serine phosphorylation
KW - Titania chromatography
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U2 - 10.1016/j.bbagen.2008.01.018
DO - 10.1016/j.bbagen.2008.01.018
M3 - Article
C2 - 18302943
AN - SCOPUS:40949158759
SN - 0304-4165
VL - 1780
SP - 716
EP - 722
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 4
ER -