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Substrate specificity and product inhibition of different forms of fructokinases and hexokinases in developing potato tubers

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Abstract

The substrate dependence and product inhibition of three different fructokinases and three different hexokinases from growing potato (Solanum tuberosum L.) tubers was investigated. The tubers contained three specific fructokinases (FK1, FK2, FK3) which had a high affinity for fructose K m=64, 90 and 100 (μM) and effectively no activity with glucose or other hexose sugars. The affinity for ATP (K m=26, 25 and 240 μM) was at least tenfold higher than for other nucleoside triphosphates. All three fructokinases showed product inhibition by high fructose (K i=5.7, 6.0 and 21 mM) and were also inhibited by ADP competitively to ATP. Sensitivity to ADP was increased in the presence of high fructose, or fructose-6-phosphate. In certain conditions, the K i (ADP) was about threefold below the K m (ATP). All three fructokinase were also inhibited by fructose-6-phosphate acting non-competitively to fructose (K i=1.3 mM for FK2). FK1 and FK2 showed very similar kinetic properties whereas FK3, which is only present at low activities in the tuber but high activities in the leaf, had a generally lower affinity for ATP, and lower sensitivity to inhibition by ADP and fructose. The tuber also contained three hexokinases (HK1, HK2, HK3) which had a high affinity for glucose (K m=41, 130 and 35 μM) and mannose but a poor affinity for fructose (K m=11, 22 and 9 mM). All three hexokinases had a tenfold higher affinity for ATP (K m=90, 280 and 560 μM) than for other nucleoside triphosphates. HK1 and HK2 were both inhibited by ADP (K i=40 and 108 μM) acting competitively to ATP. HK1, but not HK2, was inhibited by glucose-6-phosphate, which acted non-competitively to glucose (K i=4.1 mM). HK1 and HK2 differed, in that HK1 had a narrower pH optimum, a higher affinity for its substrate, and showed inhibition by glucose-6-phosphate. The relevance of these properties for the regulation of hexose metabolism in vivo is discussed.

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Abbreviations

FK:

fructokinase

Fru6P:

fructose-6-phosphate

Glc6P:

glucose-6-phosphate

HK:

hexokinase

NTP:

nucleoside triphosphate

Pi :

inorganic phosphate

UDPGlc:

uridine-5′-diphosphoglucose

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Author information

Correspondence to Mark Stitt.

Additional information

This work was supported by the Deutsche Froschungsgemeinschaft (SFB 137). We are grateful to Professor E. Beck (Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, FRG) for providing laboratory facilities.

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Renz, A., Stitt, M. Substrate specificity and product inhibition of different forms of fructokinases and hexokinases in developing potato tubers. Planta 190, 166–175 (1993). https://doi.org/10.1007/BF00196608

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Key words

  • Fructokinase
  • Hexokinase
  • Solanum
  • Sucrose metabolism