Literature summary for 2.7.9.4 extracted from

Glaring, M.A.; Zygadlo, A.; Thorneycroft, D.; Schulz, A.; Smith, S.M.; Blennow, A.; Baunsgaard, L.
An extra-plastidial alpha-glucan, water dikinase from Arabidopsis phosphorylates amylopectin in vitro and is not necessary for transient starch degradation (2007), J. Exp. Bot., 58, 3949-3960.

Cloned(Commentary)

Cloned (Comment)	Organism
in pKGWFS7 for expression N-terminally of an (eGFP)-beta-glucoronidase fusion and Agrobacterium-mediated transformation of Arabidopsis plants, in pK7FWG2 (as full-length or 158 N-terminal amino acid fragment) for expression as eGFP-fusion and subcellular localization in onion epidermis and Arabidopsis thaliana, in pPS48uYFP as full-length, C-terminal truncations (N-terminal 610, 158, or 61 amino acids) or C-terminally fused to transit peptide of AtGWD isoform 1 for Agrobacterium-mediated expression in Nicotiana benthamiana, in pGEM-4Z (as full-length or as fusion with transit peptide of AtGWD1) for in vitro transcription, translation and chloroplast import assays	Arabidopsis thaliana

Cloned (Comment)

Organism

in pKGWFS7 for expression N-terminally of an (eGFP)-beta-glucoronidase fusion and Agrobacterium-mediated transformation of Arabidopsis plants, in pK7FWG2 (as full-length or 158 N-terminal amino acid fragment) for expression as eGFP-fusion and subcellular localization in onion epidermis and Arabidopsis thaliana, in pPS48uYFP as full-length, C-terminal truncations (N-terminal 610, 158, or 61 amino acids) or C-terminally fused to transit peptide of AtGWD isoform 1 for Agrobacterium-mediated expression in Nicotiana benthamiana, in pGEM-4Z (as full-length or as fusion with transit peptide of AtGWD1) for in vitro transcription, translation and chloroplast import assays

Arabidopsis thaliana

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cell wall	along the cell wall, N-terminal 158 and 61 amino acids (revealed by confocal laser scanning microscopy)	Arabidopsis thaliana	5618	-
additional information	extra-plastidial localisation of full-length enzyme (in vitro chloroplast import assay), lack of chloroplast transit peptide	Arabidopsis thaliana	-	-
additional information	no fluorescence signal detected for full-length enzyme or its 610 N-terminal residues fused to eYFP or eGFP	Arabidopsis thaliana	-	-
nucleus	N-terminal 158 and 61 amino acids (revealed by confocal laser scanning microscopy)	Arabidopsis thaliana	5634	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + alpha-glucan + H2O	Arabidopsis thaliana	exclusive phosphorylation of C-6 position, in vitro	AMP + phospho-alpha-glucan + phosphate	-	?
additional information	Arabidopsis thaliana	not involved in transient starch degradation, plants lacking AtGWD show similar starch and sugar content as wild-type plants	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	Q9STV0	ecotype Columbia-0, wild-type or mutant lacking AtGWD2	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
phosphoprotein	autophosphorylation	Arabidopsis thaliana

Purification (Commentary)

Purification (Comment)	Organism
by metal-affinity chromatography after expression in Saccharomyces cerevisiae with C-terminal V5- and hexa-His-tag	Arabidopsis thaliana

Source Tissue

Source Tissue	Comment	Organism	Textmining
abscission zone	floral organ, strong expression in vascular tissue just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
cauline leaf	high expression level (microarray)	Arabidopsis thaliana	-
companion cell	phloem, high expression just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
flower	strong expression in mature flower parts (microarray)	Arabidopsis thaliana	-
leaf	strong expression in vascular tissue (abaxial side of primary vascular bundle, phloem) just before onset of senescence (promoter activity), senescent leaves (microarray)	Arabidopsis thaliana	-
additional information	spatially (vascular) and temporarily (age-dependent) restricted expression pattern (promoter activity)	Arabidopsis thaliana	-
root	strong expression in vascular tissue just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
sepal	strong expression in vascular tissue just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
silique	strong expression in vascular tissue (floral organ abscission zone, dehiscence zone between silique walls, vascular tissue of silique walls) just before onset of senescence (promoter activity, microarray)	Arabidopsis thaliana	-
stamen	strong expression in vascular tissue just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
stem	strong expression in vascular tissue just before onset of senescence (promoter activity)	Arabidopsis thaliana	-
vascular tissue	leaf, stem, root, flower, silique only just before onset of senescence (promoter activity)	Arabidopsis thaliana	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.02	-	amylopectin, mean chain length: 23.5, nmol glucose-6-phosphate/mg: 0.1	Arabidopsis thaliana
0.04	-	amylopectin, pre-phosphorylated, mean chain length: 24.8, nmol glucose-6-phosphate/mg: 0.5	Arabidopsis thaliana
0.49	-	amylopectin, elongated, mean chain length: 29.7, nmol glucose-6-phosphate/mg: 0.1	Arabidopsis thaliana
0.65	-	amylopectin, elongated and pre-phosphorylated, mean chain length: 30.8, nmol glucose-6-phosphate/mg: 39.4	Arabidopsis thaliana
1.36	-	soluble potato starch	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + alpha-glucan + H2O	exclusive phosphorylation of C-6 position, in vitro	Arabidopsis thaliana	AMP + phospho-alpha-glucan + phosphate	-	?
ATP + amylopectin + H2O	1 h, 30°C, pH 7, in presence of 10 mM NH4Cl, 10 mM MgCl2, 0.5 mM dithiothreitol, 0.2 mg/ml bovine serum albumin, substrates: waxy maize amylopectin (unmodified (glycogen), elongated by phosphorylase a, pre-phosphorylated by potato glucan, water dikinase (GWD), or elongated and pre-phosphorylated) or soluble potato starch	Arabidopsis thaliana	AMP + phospho-amylopectin + phosphate	reaction stop by boiling	?
additional information	not involved in transient starch degradation, plants lacking AtGWD show similar starch and sugar content as wild-type plants	Arabidopsis thaliana	?	-	?
additional information	activity is independent of pre-phosphorylation of glucan substrate	Arabidopsis thaliana	?	-	?

Synonyms

Synonyms	Comment	Organism
alpha-glucan, water dikinase	-	Arabidopsis thaliana
AtGWD2	At4g24450	Arabidopsis thaliana