P0A7D4 (PURA_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Adenylosuccinate synthetase
Short name=AMPSase
Short name=AdSS
EC=6.3.4.4

Alternative name(s):
IMP--aspartate ligase

Gene names

Name:	purA
Synonyms:	adeK
Ordered Locus Names:	b4177, JW4135

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	432 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	Plays an important role in the de novo pathway of purine nucleotide biosynthesis. Catalyzes the first committed step in the biosynthesis of AMP from IMP By similarity. HAMAP-Rule MF_00011
Catalytic activity	GTP + IMP + L-aspartate = GDP + phosphate + N(6)-(1,2-dicarboxyethyl)-AMP. HAMAP-Rule MF_00011
Cofactor	Binds 1 magnesium ion per subunit.
Pathway	Purine metabolism; AMP biosynthesis via de novo pathway; AMP from IMP: step 1/2. HAMAP-Rule MF_00011
Subunit structure	Homodimer.
Subcellular location	Cytoplasm HAMAP-Rule MF_00011.
Sequence similarities	Belongs to the adenylosuccinate synthetase family.

Ontologies

Keywords
Biological process	Purine biosynthesis
Cellular component	Cytoplasm
Ligand	GTP-binding Magnesium Metal-binding Nucleotide-binding
Molecular function	Ligase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	'de novo' AMP biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway adenosine biosynthetic process Inferred from mutant phenotype PubMed 9115248. Source: EcoliWiki nucleobase-containing small molecule interconversion Inferred from direct assay PubMed 3038024. Source: EcoliWiki purine nucleotide biosynthetic process Inferred from mutant phenotype PubMed 17012394. Source: EcoCyc response to DNA damage stimulus Inferred from expression pattern PubMed 11967071. Source: EcoliWiki
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell membrane Inferred from direct assay PubMed 16858726. Source: UniProtKB
Molecular_function	GTP binding Inferred from electronic annotation. Source: HAMAP adenylosuccinate synthase activity Inferred from direct assay Ref.19. Source: EcoCyc magnesium ion binding Inferred from electronic annotation. Source: HAMAP
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed Ref.1 Ref.6

Chain

2 – 432

431

Adenylosuccinate synthetase HAMAP-Rule MF_00011

PRO_0000095174

Regions

Nucleotide binding

13 – 19

7

GTP HAMAP-Rule MF_00011

Nucleotide binding

41 – 43

3

GTP HAMAP-Rule MF_00011

Nucleotide binding

332 – 334

3

GTP HAMAP-Rule MF_00011

Nucleotide binding

415 – 417

3

GTP HAMAP-Rule MF_00011

Region

14 – 17

4

IMP binding HAMAP-Rule MF_00011

Region

39 – 42

4

IMP binding HAMAP-Rule MF_00011

Region

300 – 306

7

Substrate binding HAMAP-Rule MF_00011

Sites

Active site

14

1

Proton acceptor

Active site

42

1

Proton donor

Metal binding

14

1

Magnesium

Metal binding

41

1

Magnesium; via carbonyl oxygen

Binding site

130

1

IMP

Binding site

144

1

IMP; shared with dimeric partner

Binding site

225

1

IMP

Binding site

240

1

IMP

Binding site

304

1

IMP

Binding site

306

1

GTP

Experimental info

Mutagenesis

13

1

G → V: Significant reduction in activity.

Mutagenesis

16

1

G → V: Significant reduction in activity.

Mutagenesis

17

1

K → Q: Reduces catalytic efficiency by 50%.

Mutagenesis

18

1

G → V: Significant reduction in activity.

Mutagenesis

19

1

K → R: Significant reduction in activity.

Mutagenesis

20

1

I → T: Significant reduction in activity.

Mutagenesis

141

1

K → I: Total loss of activity.

Mutagenesis

144

1

R → L: Does not reduce catalytic efficiency.

Mutagenesis

148

1

R → L: Reduced activity. Ref.5

Mutagenesis

304

1

R → L: Reduces catalytic efficiency by 87%.

Sequence conflict

417

1

G → D in AAA24446. Ref.1

Secondary structure

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432

Helix Strand Turn

Details...

Beta strand

4 – 13

10

Helix

17 – 24

8

Turn

25 – 27

3

Beta strand

29 – 33

5

Beta strand

42 – 46

5

Beta strand

49 – 56

8

Helix

58 – 61

4

Beta strand

62 – 64

3

Beta strand

66 – 69

4

Helix

77 – 89

13

Helix

94 – 97

4

Beta strand

98 – 100

3

Beta strand

104 – 106

3

Helix

109 – 120

12

Helix

123 – 125

3

Beta strand

126 – 128

3

Beta strand

131 – 133

3

Helix

134 – 142

9

Helix

149 – 153

5

Helix

155 – 175

21

Helix

184 – 199

16

Helix

205 – 215

11

Beta strand

219 – 222

4

Helix

227 – 229

3

Turn

231 – 233

3

Beta strand

234 – 238

5

Helix

246 – 248

3

Helix

249 – 253

5

Helix

257 – 259

3

Beta strand

262 – 273

12

Beta strand

275 – 277

3

Helix

286 – 295

10

Turn

300 – 302

3

Beta strand

307 – 309

3

Helix

313 – 323

11

Beta strand

327 – 331

5

Helix

333 – 336

4

Beta strand

340 – 349

10

Beta strand

355 – 358

4

Helix

363 – 365

3

Helix

366 – 368

3

Beta strand

370 – 377

8

Helix

389 – 391

3

Helix

394 – 407

14

Beta strand

411 – 415

5

Beta strand

417 – 419

3

Beta strand

422 – 427

6

Turn

429 – 431

3

Sequences

	Sequence		Length	Mass (Da)
	P0A7D4 [UniParc]. Last modified January 23, 2007. Version 2. Checksum: AAA862CA0F80DA70	FASTA	432	47,345
	10 20 30 40 50 60 MGNNVVVLGT QWGDEGKGKI VDLLTERAKY VVRYQGGHNA GHTLVINGEK TVLHLIPSGI 70 80 90 100 110 120 LRENVTSIIG NGVVLSPAAL MKEMKELEDR GIPVRERLLL SEACPLILDY HVALDNAREK 130 140 150 160 170 180 ARGAKAIGTT GRGIGPAYED KVARRGLRVG DLFDKETFAE KLKEVMEYHN FQLVNYYKAE 190 200 210 220 230 240 AVDYQKVLDD TMAVADILTS MVVDVSDLLD QARQRGDFVM FEGAQGTLLD IDHGTYPYVT 250 260 270 280 290 300 SSNTTAGGVA TGSGLGPRYV DYVLGILKAY STRVGAGPFP TELFDETGEF LCKQGNEFGA 310 320 330 340 350 360 TTGRRRRTGW LDTVAVRRAV QLNSLSGFCL TKLDVLDGLK EVKLCVAYRM PDGREVTTTP 370 380 390 400 410 420 LAADDWKGVE PIYETMPGWS ESTFGVKDRS GLPQAALNYI KRIEELTGVP IDIISTGPDR 430 TETMILRDPF DA

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Nucleotide sequence and analysis of the purA gene encoding adenylosuccinate synthetase of Escherichia coli K12." Wolfe S.A., Smith J.M. J. Biol. Chem. 263:19147-19153(1988) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-11. Strain: K12.
[2]	"Analysis of the Escherichia coli genome VI: DNA sequence of the region from 92.8 through 100 minutes." Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L., Blattner F.R. Nucleic Acids Res. 23:2105-2119(1995) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[3]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[4]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Evidence for an arginine residue at the substrate binding site of Escherichia coli adenylosuccinate synthetase as studied by chemical modification and site-directed mutagenesis." Dong Q., Liu F., Myers A.M., Fromm H.J. J. Biol. Chem. 266:12228-12233(1991) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 146-148, MUTAGENESIS OF ARG-148.
[6]	"Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Link A.J., Robison K., Church G.M. Electrophoresis 18:1259-1313(1997) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-10. Strain: K12 / EMG2.
[7]	"Chemical modification of adenylosuccinate synthetase from Escherichia coli by pyridoxal 5'-phosphate. Identification of an active site lysyl residue." Dong Q., Fromm H.J. J. Biol. Chem. 265:6235-6240(1990) [PubMed] [Europe PMC] [Abstract] Cited for: ACTIVE SITE LYS-141.
[8]	"Site-directed mutagenesis of the phosphate-binding consensus sequence in Escherichia coli adenylosuccinate synthetase." Liu F., Dong Q., Fromm H.J. J. Biol. Chem. 267:2388-2392(1992) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS.
[9]	"Refined crystal structures of unligated adenylosuccinate synthetase from Escherichia coli." Silva M.M., Poland B.W., Hoffman C.R., Fromm H.J., Honzatko R.B. J. Mol. Biol. 254:431-446(1995) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS), SEQUENCE REVISION TO 417.
[10]	"Refined crystal structure of adenylosuccinate synthetase from Escherichia coli complexed with hydantocidin 5'-phosphate, GDP, HPO4(2-), Mg2+, and hadacidin." Poland B.W., Lee S.F., Subramanian M.V., Siehl D.L., Anderson R.J., Fromm H.J., Honzatko R.B. Biochemistry 35:15753-15759(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) IN COMPLEX WITH GDP; MAGNESIUM; SUBSTRATE ANALOG AND INHIBITOR.
[11]	"Refined crystal structures of guanine nucleotide complexes of adenylosuccinate synthetase from Escherichia coli." Poland B.W., Hou Z., Bruns C., Fromm H.J., Honzatko R.B. J. Biol. Chem. 271:15407-15413(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 2-431.
[12]	"Crystal structures of adenylosuccinate synthetase from Escherichia coli complexed with GDP, IMP hadacidin, NO3-, and Mg2+." Poland B.W., Fromm H.J., Honzatko R.B. J. Mol. Biol. 264:1013-1027(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH GDP; IMP; MAGNESIUM AND SUBSTRATE ANALOG.
[13]	"The mode of action and the structure of a herbicide in complex with its target: binding of activated hydantocidin to the feedback regulation site of adenylosuccinate synthetase." Fonne-Pfister R., Chemla P., Ward E., Girardet M., Kreuz K.E., Honzatko R.B., Fromm H.J., Schaer H.-P., Gruetter M.G., Cowan-Jacob S.W. Proc. Natl. Acad. Sci. U.S.A. 93:9431-9436(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.55 ANGSTROMS) OF 2-431.
[14]	"Entrapment of 6-thiophosphoryl-IMP in the active site of crystalline adenylosuccinate synthetase from Escherichia coli." Poland B.W., Bruns C., Fromm H.J., Honzatko R.B. J. Biol. Chem. 272:15200-15205(1997) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) OF 2-431.
[15]	"An enzyme-bound bisubstrate hybrid inhibitor of adenylosuccinate synthetase." Hanessian S., Lu P.P., Sanceau J.Y., Chemla P., Gohda K., Fonne-Pfister R., Prade L., Cowan-Jacob S.W. Angew. Chem. Int. Ed. Engl. 38:3159-3162(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS).
[16]	"Mechanistic implications from crystalline complexes of wild-type and mutant adenylosuccinate synthetases from Escherichia coli." Choe J.Y., Poland B.W., Fromm H.J., Honzatko R.B. Biochemistry 38:6953-6961(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF WILD-TYPE AND MUTANTS GLN-17; LEU-144 AND LEU-304 IN COMPLEX WITH GTP; IMP; MAGNESIUM AND SUBSTRATE ANALOG.
[17]	"Effectors of the stringent response target the active site of Escherichia coli adenylosuccinate synthetase." Hou Z., Cashel M., Fromm H.J., Honzatko R.B. J. Biol. Chem. 274:17505-17510(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS).
[18]	"IMP alone organizes the active site of adenylosuccinate synthetase from Escherichia coli." Hou Z., Wang W., Fromm H.J., Honzatko R.B. J. Biol. Chem. 277:5970-5976(2002) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.60 ANGSTROMS) OF 1-432.
[19]	"Cavitation as a mechanism of substrate discrimination by adenylosuccinate synthetases." Iancu C.V., Zhou Y., Borza T., Fromm H.J., Honzatko R.B. Biochemistry 45:11703-11711(2006) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 2-431.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

J04199 Genomic DNA. Translation: AAA24446.1.
U14003 Genomic DNA. Translation: AAA97073.1.
U00096 Genomic DNA. Translation: AAC77134.1.
AP009048 Genomic DNA. Translation: BAE78178.1.

PIR

AJECDS. S56402.

RefSeq

NP_418598.1. NC_000913.2.
YP_492319.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1ADE	X-ray	2.00	A/B	2-432	[»]
1ADI	X-ray	2.50	A/B	2-432	[»]
1CG0	X-ray	2.50	A	2-432	[»]
1CG1	X-ray	2.50	A	2-432	[»]
1CG3	X-ray	2.50	A	2-432	[»]
1CG4	X-ray	2.50	A	2-432	[»]
1CH8	X-ray	2.50	A	2-432	[»]
1CIB	X-ray	2.30	A	2-432	[»]
1GIM	X-ray	2.50	A	2-432	[»]
1GIN	X-ray	2.80	A	2-432	[»]
1HON	X-ray	2.30	A/B	2-431	[»]
1HOO	X-ray	2.30	A/B	2-431	[»]
1HOP	X-ray	2.30	A/B	2-431	[»]
1JUY	X-ray	2.50	A	2-432	[»]
1KJX	X-ray	2.60	A	1-432	[»]
1KKB	X-ray	2.60	A	1-432	[»]
1KKF	X-ray	2.60	A	1-432	[»]
1KSZ	X-ray	2.80	A	2-431	[»]
1NHT	X-ray	2.50	A	2-431	[»]
1QF4	X-ray	2.20	A	2-432	[»]
1QF5	X-ray	2.00	A	2-432	[»]
1SON	X-ray	2.55	A	2-431	[»]
1SOO	X-ray	2.60	A	2-431	[»]
2GCQ	X-ray	2.00	A	2-431	[»]

ProteinModelPortal

P0A7D4.

SMR

P0A7D4. Positions 2-432.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-36219N.

IntAct

P0A7D4. 2 interactions.

MINT

MINT-1286548.

STRING

511145.b4177.

2D gel databases

SWISS-2DPAGE

P0A7D4.

Proteomic databases

PaxDb

P0A7D4.

PRIDE

P0A7D4.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC77134; AAC77134; b4177.
BAE78178; BAE78178; BAE78178.

GeneID

12933702.
948695.

KEGG

ecj:Y75_p4063.
eco:b4177.

PATRIC

32123927. VBIEscCol129921_4308.

Organism-specific databases

EchoBASE

EB0783.

EcoGene

EG10790. purA.

Phylogenomic databases

eggNOG

COG0104.

HOGENOM

HOG000260959.

KO

K01939.

OMA

LDYYNFQ.

ProtClustDB

PRK01117.

Enzyme and pathway databases

BioCyc

EcoCyc:ADENYLOSUCCINATE-SYN-MONOMER.
ECOL316407:JW4135-MONOMER.
MetaCyc:ADENYLOSUCCINATE-SYN-MONOMER.

UniPathway

UPA00075; UER00335.

Gene expression databases

Genevestigator

P0A7D4.

Family and domain databases

HAMAP

MF_00011. Adenylosucc_synth.

InterPro

IPR018220. Adenylosuccinate_synthase_AS.
IPR001114. Adenylosuccinate_synthetase.
IPR027417. P-loop_NTPase.
[Graphical view]

PANTHER

PTHR11846. PTHR11846. 1 hit.

Pfam

PF00709. Adenylsucc_synt. 1 hit.
[Graphical view]

SMART

SM00788. Adenylsucc_synt. 1 hit.
[Graphical view]

SUPFAM

SSF52540. SSF52540. 1 hit.

TIGRFAMs

TIGR00184. purA. 1 hit.

PROSITE

PS01266. ADENYLOSUCCIN_SYN_1. 1 hit.
PS00513. ADENYLOSUCCIN_SYN_2. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

DrugBank

DB00131. Adenosine monophosphate.

EvolutionaryTrace

P0A7D4.

Entry information

Entry name

PURA_ECOLI

Accession

Primary (citable) accession number: P0A7D4
Secondary accession number(s): P12283, Q2M6C8

Entry history

Integrated into UniProtKB/Swiss-Prot:	June 7, 2005
Last sequence update:	January 23, 2007
Last modified:	May 29, 2013
This is version 89 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program