Phosphorylation of serine-15 of maize leaf sucrose synthase: Occurrence in vivo and possible regulatory significance

Steven C Huber, Joan L. Huber, Pao Chi Liao, Douglas A. Gage, Robert W. McMichael, Prem S. Chourey, L. Curtis Hannah, Karen Koch

Research output: Contribution to journalArticle

Abstract

Experiments were conducted to determine whether sucrose synthase (SuSy) was phosphorylated in the elongation zone of maize (Zea mays L.) leaves. The approximately 90-kD subunit of SuSy was 32P-labeled on seryl residue(s) when excised shoots were fed [32P]orthophosphate. Both isoforms of SuSy (the SS1 and SS2 proteins) were phosphorylated in vivo, and tryptic peptide-mapping analysis suggested a single, similar phosphorylation site in both proteins. A combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and automated Edman sequencing analysis unequivocally identified the phosphorylation site in the maize SS2 protein as serine-15. This site was phosphorylated in vitro by endogenous protein kinase(s) in a strictly Ca2+-dependent manner. A synthetic peptide, based on the phosphorylation site sequence, was used to identify and partially purify an endogenous Ca2+-dependent protein kinase(s) from the maize leaf elongation zone and expanding spinach leaves. Phosphorylation of SuSy in vitro selectively activates the cleavage reaction by increasing the apparent affinity of the enzyme for sucrose and UDP, suggesting that phosphorylation may be of regulatory significance. Conservation of the phosphorylation site, and the sequences surrounding it, among plant species suggests that phosphorylation of SuSy may be widespread, if not universal, in plants.

Original languageEnglish (US)
Pages (from-to)793-802
Number of pages10
JournalPlant physiology
Volume112
Issue number2
DOIs
StatePublished - Oct 1996

Fingerprint

sucrose synthase
serine
Serine
Zea mays
phosphorylation
Phosphorylation
corn
leaves
protein kinases
Protein Kinases
peptide mapping
calcium
Proteins
Peptide Mapping
Uridine Diphosphate
Spinacia oleracea
proteins
matrix-assisted laser desorption-ionization mass spectrometry
synthetic peptides
orthophosphates

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

Cite this

Huber, S. C., Huber, J. L., Liao, P. C., Gage, D. A., McMichael, R. W., Chourey, P. S., ... Koch, K. (1996). Phosphorylation of serine-15 of maize leaf sucrose synthase: Occurrence in vivo and possible regulatory significance. Plant physiology, 112(2), 793-802. https://doi.org/10.1104/pp.112.2.793

Phosphorylation of serine-15 of maize leaf sucrose synthase : Occurrence in vivo and possible regulatory significance. / Huber, Steven C; Huber, Joan L.; Liao, Pao Chi; Gage, Douglas A.; McMichael, Robert W.; Chourey, Prem S.; Hannah, L. Curtis; Koch, Karen.

In: Plant physiology, Vol. 112, No. 2, 10.1996, p. 793-802.

Research output: Contribution to journalArticle

Huber, SC, Huber, JL, Liao, PC, Gage, DA, McMichael, RW, Chourey, PS, Hannah, LC & Koch, K 1996, 'Phosphorylation of serine-15 of maize leaf sucrose synthase: Occurrence in vivo and possible regulatory significance', Plant physiology, vol. 112, no. 2, pp. 793-802. https://doi.org/10.1104/pp.112.2.793
Huber, Steven C ; Huber, Joan L. ; Liao, Pao Chi ; Gage, Douglas A. ; McMichael, Robert W. ; Chourey, Prem S. ; Hannah, L. Curtis ; Koch, Karen. / Phosphorylation of serine-15 of maize leaf sucrose synthase : Occurrence in vivo and possible regulatory significance. In: Plant physiology. 1996 ; Vol. 112, No. 2. pp. 793-802.
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