Evidence for a lactose-mediated association between two nuclear carbohydrate-binding proteins.
Nuclear proteins were extracted in 2 M NaCl from membrane-depleted nuclei isolated from HL60 cells. Extracted proteins were submitted to affinity chromatography columns containing immobilized glucose, galactose or lactose. The polypeptides present in the different eluted fractions were resolved by SDS-PAGE and were either silver stained or analysed by immunoblotting with monoclonal or polyclonal antibodies, respectively, raised against the glucose-binding protein CBP67 and the galactose-binding proteins CBP35 and L14. The results presented here show that HL60 cell nuclei contain CBP35 and a glucose-binding lectin of 70 kDa (CBP70). These data account for the previously reported binding of neoglycoproteins containing glucosyl and galactosyl residues to HL60 cell nuclei. Furthermore, the present study provides the new information that CBP35 can associate with CBP70 by interactions dependent on the binding of CBP35 to lactose, and the results of some affinity chromatography experiments strongly suggest that CBP35 and CBP70 associate by protein-protein interactions. The potential function of this lactose-mediated interaction is discussed with respect to data recently reported by others showing that CBP35 is involved in in vitro mRNA splicing and that lactose inhibits the processing of the pre-RNA substrate.
Sève
A P
AP
Laboratoire de Glycobiologie et de Reconnaissance Cellulaire, INSERM U180, UFR Biomédicale des Saints-Pères, Paris, France.
Felin
M
M
Doyennette-Moyne
M A
MA
Sahraoui
T
T
Aubery
M
M
Hubert
J
J
D000943:Antigens, Differentiation / Q000302:isolation & purification / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002467:Cell Nucleus / Q000737:chemistry; D002846:Chromatography, Affinity; D037502:Galectin 3; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007785:Lactose / Q000378:metabolism*; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011956:Receptors, Cell Surface; D014407:Tumor Cells, Cultured
D000943:Antigens, Differentiation; D002352:Carrier Proteins; D037502:Galectin 3; D009687:Nuclear Proteins; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins; D005947:Glucose; D007785:Lactose