Unique modification of human heart glycerol 3-phosphate dehydrogenase by blue agarose.
The major form of glycerol phosphate dehydrogenase in human heart (GPDH-1) is a minor form (less than 15%) in brain and other tissues and is extremely labile. After GPDH-1 was eluted from an agarose column to which Cibacron blue F3GA had been covalently linked, (a) it was no longer labile (t 1/2 at 40 degrees C changed from 1.6 min to greater than 180 min); (b) it could now be stained for activity on native gels following electro-phoresis; and (c) it now migrated with the bromphenol blue dye front. The results suggest that this stabilized form of GPDH-1 is due to the covalent binding of charged ligands from the column and that this technique may be useful for studying the molecular structure and/or the active site of GPHD-1 and possibly of other enzymes which bind to blue agarose.
D055598:Chemical Phenomena; D002621:Chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D009243:NAD; D012685:Sepharose / Q000031:analogs & derivatives*
D007527:Isoenzymes; C017842:cibacron blue-sepharose; D009243:NAD; D012685:Sepharose; D005993:Glycerolphosphate Dehydrogenase