6533b7d5fe1ef96bd1265138
RESEARCH PRODUCT
Identification and purification of a stress associated nuclear carbohydrate binding protein (Mr 33000) from rat liver by application of a new photoreactive carbohydrate probe
Heinz C. SchröderGordan LaucGordan LaucWerner E.g. MüllerMirna FlögelBärbel Diehl-seifertsubject
MalePhotochemistrymedicine.drug_classMolecular Sequence DataReceptors Cell SurfaceAsialoglycoprotein ReceptorMonoclonal antibodyBiochemistryChromatography Affinitychemistry.chemical_compoundAffinity chromatographyStress PhysiologicalLectinsmedicineAnimalsMoietyDigoxigeninAmino Acid SequenceRats WistarCarbohydrate-responsive element-binding proteinMolecular BiologyCell NucleusChromatographyLysineCarbohydrate-binding proteinCell BiologyCarbohydrateRatsCross-Linking ReagentsGlucoseLiverchemistryBiochemistryMolecular ProbesRat liverElectrophoresis Polyacrylamide GelDigoxigenindescription
A photoreactive alpha-D-glucose probe has been designed for the specific detection of carbohydrate binding proteins (CBPs). The probe consists of four parts: (i) an alpha-D-glucose moiety; (ii) the digoxigenin tag; (iii) the photoreactive cross-linker; and (iv) the lysyl-lysine backbone. After incubation with lectins in the dark, the probe is activated and cross-linked to the CBPs after being treated by several flashes. Using this method we have identified a new alpha-D-glucose CBP of M(r) = 33,000, termed CBP33, in the nuclei of rats exposed to transient immobilization stress. Monoclonal antibodies were raised against the partially purified protein and subsequently used to enrich CBP33. It was purified (2400-fold) to apparent homogeneity from a 0.6 M nuclear salt extract by two subsequent affinity chromatography steps (antibody-affinity as well as alpha-D-glucose affinity column).
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1994-12-01 | Glycoconjugate Journal |