Different properties of the mitochondrial and cytosolic hexokinases in maize roots.
After tissue homogenization, 43% of the total hexokinase activity found in maize radicles was recovered in the mitochondrial fraction and 35% was soluble, in the cytosol. The maize submitochondrial particles obtained after mitochondrial sonication retained a high hexokinase activity. The mitochondrial respiration (state 4 rate) was activated by glucose. This activation was blocked by carboxyatractyloside (0.5 mM) and by oligomycin (2 micrograms/ml). The affinities for ATP and glucose of both soluble and membrane-bound maize hexokinases are similar to those of yeast hexokinase. The Km for ATP of these different forms of hexokinase varied between 0.15 and 0.37 mM, and the Km for glucose between 0.05 and 0.13 mM. A major difference between the two maize hexokinase forms is that only the mitochondrial enzyme was strongly inhibited by ADP (Ki 0.04 mM). The soluble forms of hexokinase found both in the cytosol of maize radicles and in yeast are not inhibited by ADP. In a previous report [de Meis, Grieco and Galina (1992) FEBS Lett. 308, 197-201] it was shown that the mitochondrial F1-F0-ATPase can use glucose 6-phosphate and yeast hexokinase as an ATP regenerating system. We now show that the membrane-bound hexokinase and glucose 6-phosphate can also serve as an ATP regenerating system for the mitochondria of maize radicles provided that the ADP concentration is kept below 0.05 mM. Higher ADP concentrations inhibit the reverse reaction of the mitochondrial hexokinase.
Galina
A
A
Departamento de Bioquimica, Universidade Federal do Rio de Janeiro, Brazil.
Reis
M
M
Albuquerque
M C
MC
Puyou
A G
AG
Puyou
M T
MT
de Meis
L
L
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D003600:Cytosol / Q000201:enzymology*; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000494:pharmacology; D006593:Hexokinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology*; D003313:Zea mays / Q000201:enzymology*
D005958:Glucosephosphates; D019298:Glucose-6-Phosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006593:Hexokinase; D010100:Oxygen