Isolation and characterization of multiple forms of maize leaf sucrose‐phosphate synthase

W. Kalt‐Torres, P. S. Kerr, S. C. Huber

Research output: Contribution to journalArticle

Abstract

Sucrose‐phosphate synthase SPS; (EC 2.4.1.14) from maize (Zea mays L. cv. Pioneer 3184) leaves was partially purified and kinetically characterized. Maize SPS was activated by glucose‐6‐phosphate (G‐6‐P) due to an increase in Vmax and a decrease in the Km for UDP‐glucose. The UDP‐glucose saturation profile was biphasic; thus two Km values for UDP‐glucose were calculated. Inhibition by inorganic phosphate was observed only in the presence of G‐6‐P. Chromatography of partially purified maize leaf extracts on hydroxyapatite resolved two forms of SPS activity, which differed in their affinity for UDP‐glucose and in the degree of activation by G‐6‐P. SPS was partially purified from maize leaves that were harvested in the light and in the dark. The light enzyme had a higher specific activity than the enzyme isolated from dark harvested leaves, and this difference persisted during enzyme purification. The apparent molecular weight (Stokes radius) of the light enzyme was 547 kDa, which was greater than that of the dark enzyme (457 kDa). Light and dark SPS differed in their affinities for UDP‐glucose in the absence G‐6‐P. Both the light and the dark SPS were activated by G‐6‐P; the Km for UDP‐glucose of the light enzyme was lowered by G‐6‐P, while the Km for UDP‐glucose for the dark enzyme remained unchanged. These results suggest that light activation involves a conformational change that results in differences in maximum velocity, substrate affinities and regulation by metabolites. Chromatography of either the light or dark SPS on hydroxyapatite yielded two peaks of enzyme activity, suggesting that the occurrence of the two activity peaks was not due to an interconversion of the light and dark forms.

Original languageEnglish (US)
Pages (from-to)653-658
Number of pages6
JournalPhysiologia Plantarum
Volume70
Issue number4
DOIs
StatePublished - Aug 1987

Keywords

  • Glucose‐6‐phosphate
  • hydroxyapatite
  • inorganic phosphate
  • light activation
  • metabolic regulation

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

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