Purification of Tomato Sucrose Synthase Phosphorylated Isoforms by Fe(III)-Immobilized Metal Affinity Chromatography

Anguenot, Raphaël; Yelle, Serge; Nguyen‐Quoc, Binh

doi:10.1006/abbi.1999.1146

Cited by 39 publications

(23 citation statements)

References 31 publications

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“…Studies of SUS isozymes from expanding maize leaves, pear and tomato fruits, and mung bean seedlings indicated that phosphorylation at their conserved N-terminal seryl residue activates the enzyme by increasing its affinity for sucrose and UDP (17,(27)(28)(29)31). However, the activity of soybean nodule SUS (to which RcSUS1 is most closely related; Table 2) was unaffected by phosphorylation or by phosphomimetic mutagenesis of its target phospho-site (S11D) (21).…”

Section: Discussionmentioning

confidence: 99%

“…In particular, SUS is an integral component of the plasma membranelocalized cellulose synthase complex, channeling the glucosyl moiety of UDP-glucose toward cell wall production (15,25,26). In diverse plant tissues, SUS is also phosphorylated by a calcium-dependent protein kinase (CDPK) at a conserved seryl residue located near its N terminus (1,(17)(18)(19)(20)(21)(22)(27)(28)(29)(30). However, the impact of this phosphorylation event remains somewhat enigmatic because it either activates the cleavage reaction and/or possibly mediates changes in SUS partitioning between soluble and microsomal membrane fractions (17,18,21,(27)(28)(29)31).…”

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confidence: 99%

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Biochemical and Molecular Characterization of RcSUS1, a Cytosolic Sucrose Synthase Phosphorylated in Vivo at Serine 11 in Developing Castor Oil Seeds

Fedosejevs

Ying

Park

et al. 2014

Journal of Biological Chemistry

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Background:The pathways and control of oil seed sugar unloading and metabolism are not well understood. Results: The UDP-specific sucrose synthase isozyme RcSUS1 is the dominant sucrolytic enzyme of developing castor oil seeds. Conclusion: RcSUS1 expression and phosphorylation at Ser-11 are modulated by photosynthate translocated from leaves. Significance: This research may facilitate the development of effective biotechnological strategies for oil seed metabolic engineering.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Biochemical and Molecular Characterization of RcSUS1, a Cytosolic Sucrose Synthase Phosphorylated in Vivo at Serine 11 in Developing Castor Oil Seeds

Fedosejevs

Ying

Park

et al. 2014

Journal of Biological Chemistry

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“…That 2,5-DHB and CHCA are in fact able to solubilize mono-phosphopeptides from IMAC beads contradicts the majority of IMAC affinity column elution protocols, which exploit the use of high pH to disrupt the interaction between immobilized Fe 3ϩ and the bound phosphopeptides [2,8,16,17]. The pH of 2,5-DHB in a saturated aqueous solution is 2.0, whereas traditional The supernatant from (a) was removed, the beads were washed, and the remaining phosphopeptides eluted using 100 mM ammonium phosphate buffer, pH 4.6. IMAC protocols typically use the elution conditions of sodium phosphate at high pH (8 -10.5).…”

Section: Discussionmentioning

confidence: 99%

Factors governing the solubilization of phosphopeptides retained on ferric NTA IMAC beads and their analysis by MALDI TOFMS

Hart

Waterfield

Burlingame

et al. 2002

J. Am. Soc. Mass Spectrom.

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We have revisited the direct analysis experiments reported by Tomer and co-workers in the MALDI-TOFMS analysis of phosphopeptide-loaded immobilized metal ion affinity chromatography (IMAC) beads (Zhou, W.; Merrick, B. A.; Khaledi, M. G.; Tomer, K. B. J. Am. Soc. Mass Spectrom. 2000, 11, 273-282). The results described herein provide no evidence to support a laser-induced direct desorption of phosphopeptides chelated on IMAC beads. However, we have established that solubilization of mono-phosphopeptides from their immobilized Fe 3ϩ -NTA chelates does occur effectively in solutions containing certain MALDI matrices. Particularly effective is 2,5-dihydroxybenzoic acid (2,5-DHB), which apparently forms a stronger chelation complex with Fe 3ϩ -NTA than mono-phosphopeptides. With regard to the disparity observed between the low pH value of MALDI matrices (saturated 2,5-DHB aq ϳ pH 2) and the high pH values of conventional IMAC eluents (typically above pH 7), we have also investigated the influence of eluent pH on the recovery of phosphopeptides from IMAC media. Finally, we have confirmed the importance of employing ammonium dihydrogen phosphate as buffer to achieve effective liberation of mono-and all poly-phosphopeptide species from I mmobilized metal ion affinity chromatography (IMAC) has been used for a number of years for the selective enrichment of phosphopeptides from proteolytic digest mixtures containing both phosphorylated and non-phosphorylated components [1][2][3][4][5][6][7][8]. The established methods have largely relied upon the use of high-pH elution buffers to disrupt the interaction of phosphopeptides remaining bound to the metal ioncontaining IMAC media after separation of all other peptides [2][3][4][5].In addition to the usual conditions documented above for solubilization of peptides bound in their immobilized, chelated form attached to the resin, evidence has been presented which suggests that monophosphopeptides may be released directly from the resin during the MALDI process [7,9]. It was recognized that the ferric IMAC resin must bind multiple phosphorylated components as well, but that laser irradiation does not easily dissociate these components from the agarose IMAC beads [7].While it was suggested that laser desorption directly from IMAC beads occurs particularly for mono-phosphorylated peptide components, in these reports it was also questioned whether or not the IMAC analytes are still bound to the beads through their affinity interaction immediately prior to laser desorption [9,10]. Therefore, the possibility exists that "elution" from the beads into the MALDI matrix solution/crystals has in fact taken place prior to MALDI desorption and mass analysis.Of course, there would be clear advantages in having a protocol that permits the direct analysis of phosphopeptides from IMAC beads, particularly if the analyte species thus detected could be qualitatively and quantitatively representative of the components bound to the resin. However, if phosphopeptide desorption takes place directly fro...

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“…In many dicotyledonous species such as A. thaliana (Chopra et al, 1992), Solanum tuberosum (Fu and Park, 1995), Lycopersicon esculentum (Anguenot et al, 1999), Daucus carota (Sebkova et al, 1995), C. paradisi (Komatsu et al, 2002) and P. sativum (Barratt et al, 2001), two or more sucrose synthase genes have been found. In Vicia faba and G. max, however only one Sus has been identified (Arai et al, 1992).…”

Section: Sucrose Synthase (Sus)mentioning

confidence: 99%

Role of myo-inositol phosphate synthase and sucrose synthase genes in plant seed development

Abid

Silué

Muhovski

et al. 2009

Gene

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Received by A.J. van WijnenKeywords: myo-inositol phosphate synthase (MIPS) Sucrose synthase (Sus) Seed developmentThe aim of this review is to highlight the role of myo-inositol phosphate synthase (MIPS), which catalyses the first step in inositol biosynthesis and of sucrose synthase (Sus), an enzyme involved in UDP-glucose formation, the principal nucleoside diphosphate in the sucrose cleavage reaction and in trehalose biosynthesis. These two enzymes are involved in various physiological processes including seed growth and resistance to biotic and abiotic stresses. The study of mutated MIPS and Sus genes in some crops, such as soybean and cotton, has shown that these two proteins are directly involved in embryogenesis. They exhibit several isoforms that are essential for normal seed development. The possible role of both genes in seed development is discussed in this review.

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Purification of Tomato Sucrose Synthase Phosphorylated Isoforms by Fe(III)-Immobilized Metal Affinity Chromatography

Cited by 39 publications

References 31 publications

Biochemical and Molecular Characterization of RcSUS1, a Cytosolic Sucrose Synthase Phosphorylated in Vivo at Serine 11 in Developing Castor Oil Seeds

Biochemical and Molecular Characterization of RcSUS1, a Cytosolic Sucrose Synthase Phosphorylated in Vivo at Serine 11 in Developing Castor Oil Seeds

Factors governing the solubilization of phosphopeptides retained on ferric NTA IMAC beads and their analysis by MALDI TOFMS

Role of myo-inositol phosphate synthase and sucrose synthase genes in plant seed development

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