Mass spectrometric quantification of the relative amounts of C6 and C3 position phosphorylated glucosyl residues in starch

Haebel, Sophie; Hejazi, Mahdi; Frohberg, Claus; Heydenreich, Matthias; Ritte, Gerhard

doi:10.1016/j.ab.2008.04.002

Cited by 18 publications

(18 citation statements)

References 16 publications

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“…The ratio of C3-to C6-bound phosphate in wild-type starch was ;1:5 ( Figure 6, Table 3), consistent with previous reports (Ritte et al, 2006;Haebel et al, 2008). We analyzed pwd, sex1, and sex4 starches as controls.…”

Section: Lsf2 Starch Contains High Levels Of C3-bound Phosphatesupporting

confidence: 76%

The Phosphoglucan Phosphatase Like Sex Four2 Dephosphorylates Starch at the C3-Position in Arabidopsis

et al. 2011

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Starch contains phosphate covalently bound to the C6-position (70 to 80% of total bound phosphate) and the C3-position (20 to 30%) of the glucosyl residues of the amylopectin fraction. In plants, the transient phosphorylation of starch renders the granule surface more accessible to glucan hydrolyzing enzymes and is required for proper starch degradation. Phosphate also confers desired properties to starch-derived pastes for industrial applications. In Arabidopsis thaliana, the removal of phosphate by the glucan phosphatase Starch Excess4 (SEX4) is essential for starch breakdown. We identified a homolog of SEX4, LSF2 (Like Sex Four2), as a novel enzyme involved in starch metabolism in Arabidopsis chloroplasts. Unlike SEX4, LSF2 does not have a carbohydrate binding module. Nevertheless, it binds to starch and specifically hydrolyzes phosphate from the C3-position. As a consequence, lsf2 mutant starch has elevated levels of C3-bound phosphate. SEX4 can release phosphate from both the C6-and the C3-positions, resulting in partial functional overlap with LSF2. However, compared with sex4 single mutants, the lsf2 sex4 double mutants have a more severe starch-excess phenotype, impaired growth, and a further change in the proportion of C3-and C6-bound phosphate. These findings significantly advance our understanding of the metabolism of phosphate in starch and provide innovative options for tailoring novel starches with improved functionality for industry.

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Section: Lsf2 Starch Contains High Levels Of C3-bound Phosphatesupporting

confidence: 76%

The Phosphoglucan Phosphatase Like Sex Four2 Dephosphorylates Starch at the C3-Position in Arabidopsis

et al. 2011

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“…The C-3 and 6 monophosphate peak assignments were confirmed by the spectrum of potato phosphodextrin (Fig. 2), which is known to contain monophosphate esters at C-3 and C-6 on its D-glucose repeat units (Blennow, Bay-Schmidt, Olsen, & Moller, 1998;Haebel, Hejazi, Frohberg, Heydenreich, & Ritte, 2008). The proportions of phosphate bound to the C-6 and C-3 positions of the D-glucose residues in potato starch is approximately 70-80% and 20-30%, respectively, which agrees with the intensities of the peaks in Fig.…”

Section: Resultsmentioning

confidence: 93%

“…Glucan water dikinase catalyses phosphorylation at the C-6 position from ATP, and subsequently, C-3 can be phosphorylated by phosphoglucan water dikinase Table 1 Total phosphorus contents of phosphorylated wheat starch (PWS) and of PWS after stirring for 4 h at 40°C in aqueous sodium hydroxide at pH 9.0, 10.0, 11.0, and 12.0. (Haebel et al, 2008). C-6 bound phosphate was found to have a smaller effect on helix packing than C-3 bound phosphate.…”

Section: Resultsmentioning

confidence: 94%

Effects of alkaline treatment on the structure of phosphorylated wheat starch and its digestibility

et al. 2010

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“…Samples were neutralized with 0.7 N NaOH and starch content (glucose equivalents) was quantified using the starch kit. Glucose 6-phosphate was measured as described by Haebel et al (2008). Leaf starch content was determined according to Fettke et al (2011).…”

Section: Estimation Of Starch Content and Glucose 6-phosphatementioning

confidence: 99%

Phosphorylation of transitory starch by α‐glucan, water dikinase during starch turnover affects the surface properties and morphology of starch granules

et al. 2014

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SummaryGlucan, water dikinase (GWD) is a key enzyme of starch metabolism but the physico-chemical properties of starches isolated from GWD-deficient plants and their implications for starch metabolism have so far not been described.Transgenic Arabidopsis thaliana plants with reduced or no GWD activity were used to investigate the properties of starch granules. In addition, using various in vitro assays, the action of recombinant GWD, b-amylase, isoamylase and starch synthase 1 on the surface of native starch granules was analysed.The internal structure of granules isolated from GWD mutant plants is unaffected, as thermal stability, allomorph, chain length distribution and density of starch granules were similar to wild-type. However, short glucan chain residues located at the granule surface dominate in starches of transgenic plants and impede GWD activity. A similarly reduced rate of phosphorylation by GWD was also observed in potato tuber starch fractions that differ in the proportion of accessible glucan chain residues at the granule surface.A model is proposed to explain the characteristic morphology of starch granules observed in GWD transgenic plants. The model postulates that the occupancy rate of single glucan chains at the granule surface limits accessibility to starch-related enzymes.

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Mass spectrometric quantification of the relative amounts of C6 and C3 position phosphorylated glucosyl residues in starch

Cited by 18 publications

References 16 publications

The Phosphoglucan Phosphatase Like Sex Four2 Dephosphorylates Starch at the C3-Position in Arabidopsis

The Phosphoglucan Phosphatase Like Sex Four2 Dephosphorylates Starch at the C3-Position in Arabidopsis

Effects of alkaline treatment on the structure of phosphorylated wheat starch and its digestibility

Phosphorylation of transitory starch by α‐glucan, water dikinase during starch turnover affects the surface properties and morphology of starch granules

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