Effects of activators on chemically modified yeast hexokinase.
Enzymic studies performed with chemically modified yeast hexokinase (ATP : D-hexose-6-phosphotransferase) confirm previous results indicating that the sulfhydryl, imidazol and most of the reactive amino groups do not seem to be directly implicated in the enzyme active site. On the other hand the modification of these functional groups of the enzyme does not affect the transition between the acidic inactive form to an active enzyme form after deprotonation. The chemically modified forms of hexokinase and the native enzyme are affected in the same way by activators (citrate, D-malate, 3-phosphoglycerate and Pi) when the activity was measured at pH 6.6. Moreover the loss of enzyme activity observed in the course of the chemical modifications is accompanied by an increase of the activation effect. This increase must be related to some reorganization of the enzyme active site in presence of the effectors, since the same effect was observed when hexokinase was denatured with 3M urea at pH 7.5. However no increase in the activation effect was observed when the denaturation was carried out at pH 6.5 At this pH the loss in activity and the change of optical absorption at 286 nm were much slower than at pH 7.5, which indicates a great difference in the protein structure between these pHs.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000812:Anhydrides; D001665:Binding Sites; D002951:Citrates / Q000494:pharmacology; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D005561:Formates / Q000494:pharmacology; D005988:Glyceric Acids / Q000494:pharmacology; D006593:Hexokinase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium; D008293:Malates / Q000494:pharmacology; D008751:Methylene Blue; D010710:Phosphates / Q000494:pharmacology; D010777:Photochemistry; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
D000812:Anhydrides; D002951:Citrates; D005561:Formates; D005988:Glyceric Acids; D008293:Malates; D010710:Phosphates; D000255:Adenosine Triphosphate; D014302:Trinitrobenzenesulfonic Acid; D014508:Urea; D004228:Dithionitrobenzoic Acid; D006593:Hexokinase; D008274:Magnesium; D008751:Methylene Blue