Reviewed, UniProtKB/Swiss-Prot P08179 (PUR3_ECOLI)

Names and origin

Protein names

Recommended name:
    Phosphoribosylglycinamide formyltransferase
    EC=2.1.2.2
Alternative name(s):
    5'-phosphoribosylglycinamide transformylase
    GAR transformylase
      Short name=GART

Gene names

Name:	purN
Ordered Locus Names:	b2500, JW2485

Organism

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

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

Protein attributes

Sequence length	212 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Catalytic activity	10-formyltetrahydrofolate + N(1)-(5-phospho-D-ribosyl)glycinamide = tetrahydrofolate + N(2)-formyl-N(1)-(5-phospho-D-ribosyl)glycinamide.
Pathway	Purine metabolism; IMP biosynthesis via de novo pathway; N(2)-formyl-N(1)-(5-phospho-D-ribosyl)glycinamide from N(1)-(5-phospho-D-ribosyl)glycinamide (10-formyl THF route): step 1/1.
Subunit structure	Monomer. Homodimer below pH 6.8. Ref.11
Sequence similarities	Belongs to the GART family.

Ontologies

Keywords
Biological process	Purine biosynthesis
Molecular function	Transferase
Technical term	3D-structure Complete proteome Direct protein sequencing
Gene Ontology (GO)
Biological process	'de novo' IMP biosynthetic process Inferred from electronic annotation. Source: InterPro biosynthetic process Inferred from electronic annotation. Source: InterPro
Molecular function	phosphoribosylglycinamide formyltransferase 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

Chain

1 – 212

212

Phosphoribosylglycinamide formyltransferase

PRO_0000074943

Regions

Region

11 – 13

3

5'-phosphoribosylglycinamide binding

Region

89 – 92

4

10-formyltetrahydrofolate binding

Region

140 – 144

5

10-formyltetrahydrofolate binding

Region

170 – 173

4

5'-phosphoribosylglycinamide binding

Sites

Active site

108

1

Proton donor Ref.6 Ref.8

Binding site

64

1

10-formyltetrahydrofolate

Binding site

106

1

10-formyltetrahydrofolate

Site

144

1

Raises pKa of active site His

Experimental info

Mutagenesis

70

1

E → A: Loss of homodimerization. No effect on activity.

Mutagenesis

106

1

N → A, D, G or S: Reduces activity about 2000-fold.

Mutagenesis

106

1

N → E, H, I, K, L or Y: Loss of activity.

Mutagenesis

108

1

H → A, G, M, N, Q or R: Loss of activity.

Mutagenesis

108

1

H → E, S or T: Reduces activity about 1000-fold.

Mutagenesis

119

1

H → A: No effect.

Mutagenesis

121

1

H → Q: Increases Km for 5'-phosphoribosylglycinamide 4-fold.

Mutagenesis

135

1

S → A or L: Reduces activity about 1000-fold.

Mutagenesis

137

1

H → F or Q: No effect.

Mutagenesis

144

1

D → A, E, S or Y: Reduces activity about 1000-fold.

Mutagenesis

144

1

D → C, F, H, K, L, N, P, Q, R, T or V: Loss of activity.

Secondary structure

1

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212

Helix Strand Turn

Details...

Beta strand

2 – 9

8

Helix

12 – 22

11

Beta strand

25 – 37

13

Helix

41 – 48

8

Beta strand

52 – 55

4

Helix

58 – 60

3

Beta strand

61 – 63

3

Helix

64 – 75

12

Helix

76 – 78

3

Beta strand

81 – 87

7

Helix

94 – 99

6

Turn

100 – 102

3

Beta strand

103 – 110

8

Helix

120 – 126

7

Beta strand

130 – 138

9

Turn

143 – 145

3

Beta strand

148 – 155

8

Helix

162 – 185

24

Beta strand

189 – 192

4

Beta strand

195 – 198

4

Sequences

	Sequence		Length	Mass (Da)
	P08179-1 [UniParc]. Last modified August 1, 1988. Version 1. Checksum: D38F326BCBA103FB	FASTA	212	23,238
	10 20 30 40 50 60 MNIVVLISGN GSNLQAIIDA CKTNKIKGTV RAVFSNKADA FGLERARQAG IATHTLIASA 70 80 90 100 110 120 FDSREAYDRE LIHEIDMYAP DVVVLAGFMR ILSPAFVSHY AGRLLNIHPS LLPKYPGLHT 130 140 150 160 170 180 HRQALENGDE EHGTSVHFVT DELDGGPVIL QAKVPVFAGD SEDDITARVQ TQEHAIYPLV 190 200 210 ISWFADGRLK MHENAAWLDG QRLPPQGYAA DE

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Identification and nucleotide sequence of a gene encoding 5'-phosphoribosylglycinamide transformylase in Escherichia coli K12." Smith J.M., Daum H.A. III J. Biol. Chem. 262:10565-10569(1987) [PubMed: 3301838] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[2]	"Construction of a contiguous 874-kb sequence of the Escherichia coli-K12 genome corresponding to 50.0-68.8 min on the linkage map and analysis of its sequence features." Yamamoto Y., Aiba H., Baba T., Hayashi K., Inada T., Isono K., Itoh T., Kimura S., Kitagawa M., Makino K., Miki T., Mitsuhashi N., Mizobuchi K., Mori H., Nakade S., Nakamura Y., Nashimoto H., Oshima T. , Oyama S., Saito N., Sampei G., Satoh Y., Sivasundaram S., Tagami H., Takahashi H., Takeda J., Takemoto K., Uehara K., Wada C., Yamagata S., Horiuchi T. DNA Res. 4:91-113(1997) [PubMed: 9205837] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[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: 9278503] [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: 16738553] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Multiple independent origins of Shigella clones of Escherichia coli and convergent evolution of many of their characteristics." Pupo G.M., Lan R., Reeves P.R. Proc. Natl. Acad. Sci. U.S.A. 97:10567-10572(2000) [PubMed: 10954745] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: ECOR 7.
[6]	"Active-site mapping and site-specific mutagenesis of glycinamide ribonucleotide transformylase from Escherichia coli." Inglese J., Smith J.M., Benkovic S.J. Biochemistry 29:6678-6687(1990) [PubMed: 2204419] [Abstract] Cited for: ACTIVE SITE, PARTIAL PROTEIN SEQUENCE.
[7]	"A rapid screen of active site mutants in glycinamide ribonucleotide transformylase." Warren M.S., Marolewski A.E., Benkovic S.J. Biochemistry 35:8855-8862(1996) [PubMed: 8688421] [Abstract] Cited for: MUTAGENESIS.
[8]	"Catalytic mechanism of Escherichia coli glycinamide ribonucleotide transformylase probed by site-directed mutagenesis and pH-dependent studies." Shim J.H., Benkovic S.J. Biochemistry 38:10024-10031(1999) [PubMed: 10433709] [Abstract] Cited for: MUTAGENESIS, ACTIVE SITE.
[9]	"Crystal structure of glycinamide ribonucleotide transformylase from Escherichia coli at 3.0-A resolution. A target enzyme for chemotherapy." Chen P., Schulze-Gahmen U., Stura E.A., Inglese J., Johnson D.L., Marolewski A., Benkovic S.J., Wilson I.A. J. Mol. Biol. 227:283-292(1992) [PubMed: 1522592] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (3.0 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG.
[10]	"Structures of apo and complexed Escherichia coli glycinamide ribonucleotide transformylase." Almassy R.J., Janson C.A., Kan C.-C., Hostomska Z. Proc. Natl. Acad. Sci. U.S.A. 89:6114-6118(1992) [PubMed: 1631098] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS.
[11]	"A pH-dependent stabilization of an active site loop observed from low and high pH crystal structures of mutant monomeric glycinamide ribonucleotide transformylase at 1.8 to 1.9 A." Su Y., Yamashita M.M., Greasley S.E., Mullen C.A., Shim J.H., Jennings P.A., Benkovic S.J., Wilson I.A. J. Mol. Biol. 281:485-499(1998) [PubMed: 9698564] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF MUTANT ALA-70 IN COMPLEX WITH SUBSTRATE ANALOGS, SUBUNIT.
[12]	"New insights into inhibitor design from the crystal structure and NMR studies of Escherichia coli GAR transformylase in complex with beta-GAR and 10-formyl-5,8,10-trideazafolic acid." Greasley S.E., Yamashita M.M., Cai H., Benkovic S.J., Boger D.L., Wilson I.A. Biochemistry 38:16783-16793(1999) [PubMed: 10606510] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

M13747 Genomic DNA. Translation: AAA83899.1.
U00096 Genomic DNA. Translation: AAC75553.1.
AP009048 Genomic DNA. Translation: BAA16388.1.
AF293167 Genomic DNA. Translation: AAG14584.1.

PIR

XYECGF. A28486.

RefSeq

AP_003086.1.
NP_416995.1.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1C2T	X-ray	2.10	A/B	1-212	[»]
1C3E	X-ray	2.10	A/B	1-209	[»]
1CDD	X-ray	2.80	A/B	1-212	[»]
1CDE	X-ray	2.50	A/B/C/D	1-212	[»]
1GAR	X-ray	1.96	A/B	1-212	[»]
1GRC	X-ray	3.00	A/B	1-212	[»]
1JKX	X-ray	1.60	A/B/C/D	1-212	[»]
2GAR	X-ray	1.80	A	1-212	[»]
3GAR	X-ray	1.90	A	1-212	[»]

ModBase

Search...

Genome annotation databases

GeneID

946973.

GenomeReviews

Gene locus JW2485 in contig AP009048_GR.
Gene locus b2500 in contig U00096_GR.

KEGG

ecj:JW2485.
eco:b2500.

Organism-specific databases

EchoBASE

EB0792.

EcoGene

EG10799. purN.

CMR

Search...

Phylogenomic databases

HOGENOM

P08179.

OMA

P08179. LASWRGT.

Enzyme and pathway databases

BioCyc

EcoCyc:GART-MON.
MetaCyc:GART-MON.

Family and domain databases

InterPro

IPR002376. Formyl_transf_N.
IPR001555. GART_AS.
IPR004607. PurN_trans.
[Graphical view]

Gene3D

G3DSA:3.40.50.170. Formyl_transf_N. 1 hit.

Pfam

PF00551. Formyl_trans_N. 1 hit.
[Graphical view]

TIGRFAMs

TIGR00639. PurN. 1 hit.

PROSITE

PS00373. GART. 1 hit.
[Graphical view]

ProtoNet

Search...

Other Resources

BindingDB

P08179.

Entry information

Entry name

PUR3_ECOLI

Accession

Primary (citable) accession number: P08179

Entry history

Integrated into UniProtKB/Swiss-Prot:	August 1, 1988
Last sequence update:	August 1, 1988
Last modified:	June 16, 2009
This is version 96 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)