Q9AWA5 (GWD1_SOLTU) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Alpha-glucan water dikinase, chloroplastic
EC=2.7.13.3
EC=2.7.9.4

Alternative name(s):
Starch-related R1 protein

Gene names

Name:

R1

Organism

Solanum tuberosum (Potato)

Taxonomic identifier

4113 [NCBI]

Taxonomic lineage

Eukaryota › Viridiplantae › Streptophyta › Embryophyta › Tracheophyta › Spermatophyta › Magnoliophyta › eudicotyledons › core eudicotyledons › asterids › lamiids › Solanales › Solanaceae › Solanoideae › Solaneae › Solanum

Protein attributes

Sequence length	1464 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Mediates the incorporation of phosphate into starch-like alpha-glucan, mostly at the C-6 position of glucose units. Acts as an overall regulator of starch mobilization. Required for starch degradation, suggesting that the phosphate content of starch regulates its degradability. More active on alpha-1,6 branched amylopectin. Ref.1 Ref.2 Ref.6
Catalytic activity	ATP + alpha-glucan + H2O = AMP + phospho-alpha-glucan + phosphate. Ref.5 ATP + protein L-histidine = ADP + protein N-phospho-L-histidine. Ref.5
Cofactor	Magnesium. Ref.5 Magnesium By similarity. Ref.5
Subunit structure	Homodimer. Ref.6
Subcellular location	Plastid › chloroplast. Note: Starch granules. Binds to starch granules isolated from either illuminated or darkened leaves. However, binding to darkened leaves is more effective, suggesting that depending upon the metabolic state of the cells, the starch granule surface changes and thereby affects its binding. Ref.1 Ref.4
Tissue specificity	Expressed in leaves. Ref.1
Domain	The N-terminal domain contains the alpha-glucan binding site, the central domain the pyrophosphate/phosphate carrier histidine, and the C-terminal domain the ATP binding site.
Miscellaneous	The reaction takes place in three steps, mediated by a phosphocarrier histidine residue located on the surface of the central domain. The two first partial reactions are catalyzed at an active site located on the C-terminal domain, and the third partial reaction is catalyzed at an active site located on the N-terminal domain. For catalytic turnover, the central domain swivels from the concave surface of the C-terminal domain to that of the N-terminal domain.
Sequence similarities	Belongs to the PEP-utilizing enzyme family.
Biophysicochemical properties	Kinetic parameters: KM=0.23 µM for ATP Ref.5 Ref.6 KM=1.7 µM for amylopectin pH dependence: Optimum pH is 7. Temperature dependence: Optimum temperature is 35 degrees Celsius.

Ontologies

Keywords
Biological process	Carbohydrate metabolism
Cellular component	Chloroplast Plastid
Domain	Transit peptide
Ligand	ATP-binding Magnesium Metal-binding Nucleotide-binding
Molecular function	Kinase Transferase
Technical term	Direct protein sequencing
Gene Ontology (GO)
Biological process	carbohydrate metabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	chloroplast Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW alpha-glucan, water dikinase activity Inferred from electronic annotation. Source: EC ligase activity Inferred from electronic annotation. Source: InterPro metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW protein histidine kinase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Transit peptide	1 – 77	77	Chloroplast Ref.1 Ref.3
	Chain	78 – 1464	1387	Alpha-glucan water dikinase, chloroplastic		PRO_0000023567
Sites
	Active site	1069	1	Tele-phosphohistidine intermediate Ref.3 Ref.6
Experimental info
	Mutagenesis	1069	1	H → A: Loss of activity, no autophosphorylation. Ref.6
	Sequence conflict	66	1	E → G Ref.2
	Sequence conflict	68	1	K → N Ref.2
	Sequence conflict	86	1	E → Q in AAK11735. Ref.2
	Sequence conflict	86	1	E → Q AA sequence Ref.3
	Sequence conflict	94	1	G → E in AAK11735. Ref.2
	Sequence conflict	198	1	N → I in AAK11735. Ref.2
	Sequence conflict	203 – 204	2	RV → HI in AAK11735. Ref.2
	Sequence conflict	240	1	P → T in AAK11735. Ref.2
	Sequence conflict	254	1	V → E in AAK11735. Ref.2
	Sequence conflict	292	1	D → N in AAK11735. Ref.2
	Sequence conflict	346	1	T → K in AAK11735. Ref.2
	Sequence conflict	353	1	K → E in AAK11735. Ref.2
	Sequence conflict	379	1	H → Q in AAK11735. Ref.2
	Sequence conflict	385	1	T → P in AAK11735. Ref.2
	Sequence conflict	400	1	P → A in AAK11735. Ref.2
	Sequence conflict	400	1	Missing AA sequence Ref.3
	Sequence conflict	438	1	A → S in AAK11735. Ref.2
	Sequence conflict	540	1	G → D in AAK11735. Ref.2
	Sequence conflict	547	1	L → S in AAK11735. Ref.2
	Sequence conflict	588	1	I → M in AAK11735. Ref.2
	Sequence conflict	681	1	N → K in AAK11735. Ref.2
	Sequence conflict	690	1	Q → E in AAK11735. Ref.2
	Sequence conflict	768	1	R → K in AAK11735. Ref.2
	Sequence conflict	1129	1	A → V in AAK11735. Ref.2
	Sequence conflict	1129	1	A → V AA sequence Ref.3
	Sequence conflict	1209	1	M → T in AAK11735. Ref.2
	Sequence conflict	1256	1	G → V in AAK11735. Ref.2

Sequences

	Sequence		Length	Mass (Da)
	Q9AWA5 [UniParc]. Last modified January 4, 2005. Version 2. Checksum: 62B0FE0FE298EE24	FASTA	1,464	163,237
	10 20 30 40 50 60 MSNSLGNNLL YQGFLTSTVL EHKSRISPPC VGGNSLFQQQ VISKSPLSTE FRGNRLKVQK 70 80 90 100 110 120 KKIPMEKKRA FSSSPHAVLT TDTSSELAEK FSLGGNIELQ VDVRPPTSGD VSFVDFQVTN 130 140 150 160 170 180 GSDKLFLHWG AVKFGKETWS LPNDRPDGTK VYKNKALRTP FVKSGSNSIL RLEIRDTAIE 190 200 210 220 230 240 AIEFLIYDEA HDKWIKNNGG NFRVKLSRKE IRGPDVSVPE ELVQIQSYLR WERKGKQNYP 250 260 270 280 290 300 PEKEKEEYEA ARTVLQEEIA RGASIQDIRA RLTKTNDKSQ SKEEPLHVTK SDIPDDLAQA 310 320 330 340 350 360 QAYIRWEKAG KPNYPPEKQI EELEEARREL QLELEKGITL DELRKTITKG EIKTKVEKHL 370 380 390 400 410 420 KRSSFAVERI QRKKRDFGHL INKYTSSPAV QVQKVLEEPP ALSKIKLYAK EKEEQIDDPI 430 440 450 460 470 480 LNKKIFKVDD GELLVLVAKS SGKTKVHLAT DLNQPITLHW ALSKSPGEWM VPPSSILPPG 490 500 510 520 530 540 SIILDKAAET PFSASSSDGL TSKVQSLDIV IEDGNFVGMP FVLLSGEKWI KNQGSDFYVG 550 560 570 580 590 600 FSAASKLALK AAGDGSGTAK SLLDKIADME SEAQKSFMHR FNIAADLIED ATSAGELGFA 610 620 630 640 650 660 GILVWMRFMA TRQLIWNKNY NVKPREISKA QDRLTDLLQN AFTSHPQYRE ILRMIMSTVG 670 680 690 700 710 720 RGGEGDVGQR IRDEILVIQR NNDCKGGMMQ EWHQKLHNNT SPDDVVICQA LIDYIKSDFD 730 740 750 760 770 780 LGVYWKTLNE NGITKERLLS YDRAIHSEPN FRGDQKGGLL RDLGHYMRTL KAVHSGADLE 790 800 810 820 830 840 SAIANCMGYK TEGEGFMVGV QINPVSGLPS GFQDLLHFVL DHVEDKNVET LLERLLEARE 850 860 870 880 890 900 ELRPLLLKPN NRLKDLLFLD IALDSTVRTA VERGYEELNN ANPEKIMYFI SLVLENLALS 910 920 930 940 950 960 VDDNEDLVYC LKGWNQALSM SNGGDNHWAL FAKAVLDRTR LALASKAEWY HHLLQPSAEY 970 980 990 1000 1010 1020 LGSILGVDQW ALNIFTEEII RAGSAASLSS LLNRLDPVLR KTANLGSWQI ISPVEAVGYV 1030 1040 1050 1060 1070 1080 VVVDELLSVQ NEIYEKPTIL VAKSVKGEEE IPDGAVALIT PDMPDVLSHV SVRARNGKVC 1090 1100 1110 1120 1130 1140 FATCFDPNIL ADLQAKEGRI LLLKPTPSDI IYSEVNEIEL QSSSNLVEAE TSATLRLVKK 1150 1160 1170 1180 1190 1200 QFGGCYAISA DEFTSEMVGA KSRNIAYLKG KVPSSVGIPT SVALPFGVFE KVLSDDINQG 1210 1220 1230 1240 1250 1260 VAKELQILMK KLSEGDFSAL GEIRTTVLDL SAPAQLVKEL KEKMQGSGMP WPGDEGPKRW 1270 1280 1290 1300 1310 1320 EQAWMAIKKV WASKWNERAY FSTRKVKLDH DYLCMAVLVQ EIINADYAFV IHTTNPSSGD 1330 1340 1350 1360 1370 1380 DSEIYAEVVR GLGETLVGAY PGRALSFICK KKDLNSPQVL GYPSKPIGLF IKRSIIFRSD 1390 1400 1410 1420 1430 1440 SNGEDLEGYA GAGLYDSVPM DEEEKVVIDY SSDPLITDGN FRQTILSNIA RAGHAIEELY 1450 1460 GSPQDIEGVV RDGKIYVVQT RPQM

References

[1]	"Inhibition of a starch-granule-bound protein leads to modified starch and repression of cold sweetening." Lorberth R., Ritte G., Willmitzer L., Kossmann J. Nat. Biotechnol. 16:473-477(1998) [PubMed: 9592398] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 78-91, FUNCTION, SUBCELLULAR LOCATION, TISSUE SPECIFICITY. Strain: cv. Desiree.
[2]	"Production of highly phosphorylated glycopolymers by expression of R1 in Escherichia coli." Viksoe-Nielsen A., Chen P.-H.J., Larsson H., Blennow A., Moeller B.L. Carbohydr. Res. 337:327-333(2002) [PubMed: 11841813] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], FUNCTION. Strain: cv. Prevalent.
[3]	"Functional domain organization of the potato alpha-glucan, water dikinase (GWD): evidence for separate site catalysis as revealed by limited proteolysis and deletion mutants." Mikkelsen R., Blennow A. Biochem. J. 385:355-361(2005) [PubMed: 15361065] [Abstract] Cited for: PROTEIN SEQUENCE OF 78-86; 394-405; 614-622; 797-805; 1121-1130 AND 1132-1140, ACTIVE SITE HIS-1069, ENZYMIC MECHANISM.
[4]	"Reversible binding of the starch-related R1 protein to the surface of transitory starch granules." Ritte G., Lorberth R., Steup M. Plant J. 21:387-391(2000) [PubMed: 10758490] [Abstract] Cited for: SUBCELLULAR LOCATION.
[5]	"The starch-related R1 protein is an alpha-glucan, water dikinase." Ritte G., Lloyd J.R., Eckermann N., Rottmann A., Kossmann J., Steup M. Proc. Natl. Acad. Sci. U.S.A. 99:7166-7171(2002) [PubMed: 12011472] [Abstract] Cited for: ENZYME ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES.
[6]	"Functional characterization of alpha-glucan, water dikinase, the starch phosphorylating enzyme." Mikkelsen R., Baunsgaard L., Blennow A. Biochem. J. 377:525-532(2004) [PubMed: 14525539] [Abstract] Cited for: FUNCTION, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE HIS-1069, MUTAGENESIS OF HIS-1069.

Cross-references

Sequence databases
EMBL GenBank DDBJ	Y09533 mRNA. Translation: CAA70725.1. AY027522 mRNA. Translation: AAK11735.1.
PIR	T07050.
3D structure databases
ProteinModelPortal	Q9AWA5.
ModBase	Search...
Protein family/group databases
CAZy	CBM45. Carbohydrate-Binding Module Family 45.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Enzyme and pathway databases
BRENDA	2.7.9.4. 5757.
Family and domain databases
InterPro	IPR013815. ATP_grasp_subdomain_1. IPR013816. ATP_grasp_subdomain_2. IPR002192. PPDK_PEP-bd. [Graphical view]
Gene3D	G3DSA:3.30.1490.20. ATP_grasp_subdomain_1. 1 hit. G3DSA:3.30.470.20. ATP_grasp_subdomain_2. 1 hit.
Pfam	PF01326. PPDK_N. 1 hit. [Graphical view]
PROSITE	PS00742. PEP_ENZYMES_2. False negative. PS00370. PEP_ENZYMES_PHOS_SITE. False negative. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

GWD1_SOLTU

Accession

Primary (citable) accession number: Q9AWA5
Secondary accession number(s): O82061

Entry history

Integrated into UniProtKB/Swiss-Prot:	January 4, 2005
Last sequence update:	January 4, 2005
Last modified:	September 21, 2011
This is version 50 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Plant Protein Annotation Program