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

Fedosejevs, Eric T.; Ying, Sheng; Park, Joonho; Anderson, Erin M.; Mullen, Robert T.; She, Yi‐Min; Plaxton, William C.

doi:10.1074/jbc.m114.585554

Cited by 24 publications

(32 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Furthermore, RcCDPK2 phosphorylates RcSUS1 at Ser-11 in developing COS and shows many additional and prominent differences compared to RcCDPK1 including: (i) a relatively nonspecific in vitro substrate selectivity, including abundant peptide kinase activity; (ii) a significantly lower K 0.5 (Ca 2+ ) value of approximately 200 nM; (iii) insensitivity to metabolite effectors including PEP; (iv) partial association with microsomal membranes; and (iv) a completely opposite developmental profile in COS (i.e. RcCDPK2 expression, RcSUS1 Ser-11 kinase activity, and in vivo RcSUS1 phosphorylation at Ser-11 progressively decrease during COS development; Fedosejevs et al, 2014Fedosejevs et al, , 2016.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, all PTPCs and Suc synthases examined to date share an orthologous phosphorylation motif near their N-terminus, i.e. f -5 -X -4 -basic -3 -X -2 -X -1 -Ser-X +1 -X +2 -X +3 -f +4 (where f is a hydrophobic residue, X is any amino acid, and subscripts denote residue positions relative to the seryl phosphorylation site; Winter and Huber, 2000;O'Leary et al, 2011b;Fedosejevs et al, 2014Fedosejevs et al, , 2016. This differs from the unique and highly conserved BTPC recognition motif identified for RcCDPK1, namely: f -4 -X -3 -Basic -2 -X -1 -Ser-X +1 -X +2 -Basic +3 -f +4 (Dalziel et al, 2012;Hill et al, 2014;Supplemental Fig.…”

Section: Rccdpk1 Is Highly Expressed During Castor Bean Developmentmentioning

confidence: 99%

“…Results of (A) and (B) are representative of three independent experiments. Leary et al, 2009;Dalziel et al, 2012), in vitro dephosphorylated COS Class-1 PEPC or RcSUS1 (Tripodi et al, 2005;Fedosejevs et al, 2014), or histone III-S from calf thymus (10 mg) were incubated for 10 min at 30°C in 25-mL of the standard [g-32 P]ATP phosphorylation assay mix containing 0.2 mM CaCl 2 and 250 ng of RcCDPK1. After SDS-PAGE (2 mg protein per lane), the gel was subjected to (A) phosphorimaging and (B) stained with Coomassie Blue R-250 (CBB-250).…”

Section: +mentioning

confidence: 99%

See 2 more Smart Citations

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

et al. 2017

Self Cite

View full text Add to dashboard Cite

Phosphoenolpyruvate carboxylase (PEPC) is a tightly controlled cytosolic enzyme situated at a crucial branch point of central plant metabolism. In developing castor oil seeds (Ricinus communis) a novel, allosterically desensitized 910-kD Class-2 PEPC hetero-octameric complex, arises from a tight interaction between 107-kD plant-type PEPC and 118-kD bacterial-type (BTPC) subunits. The native Ca 2+ -dependent protein kinase (CDPK) responsible for in vivo inhibitory phosphorylation of Class-2 PEPC's BTPC subunit's at Ser-451 was highly purified from COS and identified as RcCDPK1 (XP_002526815) by mass spectrometry. Heterologously expressed RcCDPK1 catalyzed Ca 2+ -dependent, inhibitory phosphorylation of BTPC at Ser-451 while exhibiting: (i) a pair of Ca 2+ binding sites with identical dissociation constants of 5.03 mM, (ii) a Ca 2+ -dependent electrophoretic mobility shift, and (iii) a marked Ca 2+ -independent hydrophobicity. Pull-down experiments established the Ca 2+ -dependent interaction of N-terminal GST-tagged RcCDPK1 with BTPC. RcCDPK1-Cherry localized to the cytosol and nucleus of tobacco bright yellow-2 cells, but colocalized with mitochondrial-surface associated BTPC-enhanced yellow fluorescent protein when both fusion proteins were coexpressed. Deletion analyses demonstrated that although its N-terminal variable domain plays an essential role in optimizing Ca 2+ -dependent RcCDPK1 autophosphorylation and BTPC transphosphorylation activity, it is not critical for in vitro or in vivo target recognition. Arabidopsis (Arabidopsis thaliana) CPK4 and soybean (Glycine max) CDPKb are RcCDPK1 orthologs that effectively phosphorylated castor BTPC at Ser-451. Overall, the results highlight a potential link between cytosolic Ca 2+ signaling and the posttranslational control of respiratory CO 2 refixation and anaplerotic photosynthate partitioning in support of storage oil and protein biosynthesis in developing COS.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Rccdpk1 Is Highly Expressed During Castor Bean Developmentmentioning

confidence: 99%

Section: +mentioning

confidence: 99%

See 1 more Smart Citation

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Similarly, Suc synthase, involved in the breakdown of Suc in heterotrophic tissues (e.g. the nonphotosynthetic base of a developing maize leaf and developing seeds) is phosphorylated by a Ca 2+ -dependent protein kinase; the phosphorylation influences both protein activity and membrane association (Duncan and Huber, 2007;Fedosejevs et al, 2014).…”

Section: (Classic) Examples Of Plant Metabolic Enzymes and Pathways Rmentioning

confidence: 99%

Update: Post-translational protein modifications in plant metabolism

2015

View full text Add to dashboard Cite

ORCID ID: 0000-0001-9536-0487 (K.J.v.W.).Posttranslational modifications (PTMs) of proteins greatly expand proteome diversity, increase functionality, and allow for rapid responses, all at relatively low costs for the cell. PTMs play key roles in plants through their impact on signaling, gene expression, protein stability and interactions, and enzyme kinetics. Following a brief discussion of the experimental and bioinformatics challenges of PTM identification, localization, and quantification (occupancy), a concise overview is provided of the major PTMs and their (potential) functional consequences in plants, with emphasis on plant metabolism. Classic examples that illustrate the regulation of plant metabolic enzymes and pathways by PTMs and their cross talk are summarized. Recent large-scale proteomics studies mapped many PTMs to a wide range of metabolic functions. Unraveling of the PTM code, i.e. a predictive understanding of the (combinatorial) consequences of PTMs, is needed to convert this growing wealth of data into an understanding of plant metabolic regulation.The primary amino acid sequence of proteins is defined by the translated mRNA, often followed by Nor C-terminal cleavages for preprocessing, maturation, and/or activation. Proteins can undergo further reversible or irreversible posttranslational modifications (PTMs) of specific amino acid residues. Proteins are directly responsible for the production of plant metabolites because they act as enzymes or as regulators of enzymes. Ultimately, most proteins in a plant cell can affect plant metabolism (e.g. through effects on plant gene expression, cell fate and development, structural support, transport, etc.). Many metabolic enzymes and their regulators undergo a variety of PTMs, possibly resulting in changes in oligomeric state, stabilization/degradation, and (de)activation (Huber and Hardin, 2004), and PTMs can facilitate the optimization of metabolic flux. However, the direct in vivo consequence of a PTM on a metabolic enzyme or pathway is frequently not very clear, in part because it requires measurements of input and output of the reactions, including flux through the enzyme or pathway. This Update will start out with a short overview on the major PTMs observed for each amino acid residue (Table I), followed by a brief discussion of techniques for the characterization of protein PTMs, including determination of the localization within proteins (i.e. the specific residues) and occupancy. Challenges in dealing with multiple PTMs per protein and cross talk between PTMs will be briefly outlined. We then describe the major physiological PTMs observed in plants as well as PTMs that are nonenzymatically induced during sample preparation (Table II). Finally, this Update is completed with a number of well-studied, classical examples of the regulation of plant metabolism by PTMs, in particular for enzymes in primary metabolism (Calvin cycle, glycolysis, and respiration) and the C4 shuttle accommodating photosynthesis in C4 plants (Table III). As will be ev...

show abstract

“…No endosperma da semente, o polissacarídeo de armazenamento mais comum é o amido. Encontrado tanto na forma de amilose quanto amilopectina, é sintetizado a partir de ADP-glicose, que por sua vez, é sintetizada a partir de glicose-1-fosfato e ATP em reação catalisada pela enzima AGPase (SPIELBAUER et al, 2006;FEDOSEJEVS et al, 2014). Em plantas, esta enzima é heterotetrâmera, conformada por duas subunidades grandes e duas pequenas, que são codificadas pelos genes shrunken-2 (Sh2) e brittle-2 (Bt2), respectivamente (SIVAK, PREISS, 1994;MÉCHIN et al, 2004).…”

Section: Perfil Proteico De Variedades Gm E Cgunclassified

Estudo proteômico de variedades de milho (<i>Zea mays</i> L.) obtidas por melhoramento clássico e por recombinação genética

Santos-Donado¹

View full text Add to dashboard Cite

Estudo proteômico de variedades de milho (Zea mays L.)obtidas por melhoramento clássico e por recombinação genética SÃO PAULO 2016 Priscila Robertina dos Santos-Donado processes and molecular function. In conclusion, there were found differences between the cultures of GMs and CGs, indicating a normal variation among the corn varieties, which do not affect the food security of the studied samples. As per the samples with QPM and CN, the differences found were due to the line distances or germplasm. Estudo proteômico de variedades de milho (Zea mays L.) obtidas por melhoramento clássico e por recombinação genética

show abstract

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

Cited by 24 publications

References 59 publications

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

Update: Post-translational protein modifications in plant metabolism

Estudo proteômico de variedades de milho (<i>Zea mays</i> L.) obtidas por melhoramento clássico e por recombinação genética

Contact Info

Product

Resources

About

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

Cited by 24 publications

References 59 publications

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca2+-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca2+-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

Update: Post-translational protein modifications in plant metabolism

Estudo proteômico de variedades de milho (<i>Zea mays</i> L.) obtidas por melhoramento clássico e por recombinação genética

Contact Info

Product

Resources

About

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds

Regulatory Phosphorylation of Bacterial-Type PEP Carboxylase by the Ca²⁺-Dependent Protein Kinase RcCDPK1 in Developing Castor Oil Seeds