Reviewed, UniProtKB/Swiss-Prot P16659 (SYP_ECOLI)

Names and origin

Protein names

Recommended name:
    Prolyl-tRNA synthetase
    EC=6.1.1.15
Alternative name(s):
    Proline--tRNA ligase
      Short name=ProRS
    Global RNA synthesis factor

Gene names

Name:	proS
Synonyms:	drpA
Ordered Locus Names:	b0194, JW0190

Organism

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

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

Protein attributes

Sequence length	572 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Catalyzes the attachment of proline to tRNA(Pro) in a two-step reaction: proline is first activated by ATP to form Pro-AMP and then transferred to the acceptor end of tRNA(Pro). As ProRS can inadvertently accommodate and process non-cognate amino acids such as alanine and cysteine, to avoid such errors it has two additional distinct editing activities against alanine. One activity is designated as 'pretransfer' editing and involves the tRNA(Pro)-independent hydrolysis of activated Ala-AMP. The other activity is designated 'posttransfer' editing and involves deacylation of mischarged Ala-tRNA(Pro). The misacylated Cys-tRNA(Pro) is not edited by ProRS. HAMAP MF_01569
Catalytic activity	ATP + L-proline + tRNA(Pro) = AMP + diphosphate + L-prolyl-tRNA(Pro). HAMAP MF_01569
Subunit structure	Homodimer. HAMAP MF_01569
Subcellular location	Cytoplasm. HAMAP MF_01569
Domain	Consists of three domains: the N-terminal catalytic domain, the editing domain and the C-terminal anticodon-binding domain. Ref.13
Sequence similarities	Belongs to the class-II aminoacyl-tRNA synthetase family. ProS type 1 subfamily.
Biophysicochemical properties	Kinetic parameters: KM=0.25 mM for proline Ref.9 Ref.10 Ref.12 KM=140 mM for alanine KM=0.17 mM for cysteine

Ontologies

Keywords
Biological process	Protein biosynthesis
Cellular component	Cytoplasm
Ligand	ATP-binding Nucleotide-binding
Molecular function	Aminoacyl-tRNA synthetase Ligase
Technical term	Complete proteome Direct protein sequencing
Gene Ontology (GO)
Biological process	prolyl-tRNA aminoacylation Inferred from electronic annotation. Source: HAMAP
Cellular component	cytosol Inferred from direct assay. Source: UniProtKB
Molecular function	ATP binding Inferred from electronic annotation. Source: HAMAP proline-tRNA ligase activity Inferred from electronic annotation. Source: HAMAP protein binding Inferred from physical interaction. Source: IntAct
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
groL	P0A6F5	1	EBI-549550,EBI-543750

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 572	572	Prolyl-tRNA synthetase HAMAP MF_01569		PRO_0000139329
Experimental info
	Mutagenesis	257	1	T → A: Reduces the posttransfer editing activity 5-fold, with little change in both the aminoacylation and the pretransfer editing activities. Ref.11
	Mutagenesis	279	1	K → A: Severely affects the posttransfer editing activity, with little change in both the aminoacylation and the pretransfer editing activities. Ref.11
	Mutagenesis	350	1	D → A: Abolishes the pretransfer editing activity and reduces the aminoacylation activity 20-fold and the posttransfer editing activity 5-fold. Ref.11
	Mutagenesis	369	1	H → A: Reduces both the aminoacylation and the pretransfer editing activities 2.5-fold, and the posttransfer editing activity 5-fold. Loss of specificity in deacylation. Ref.11
	Mutagenesis	378	1	D → A: Little change in both the aminoacylation and the editing activities. Ref.11
	Mutagenesis	386	1	D → A: Reduces the posttransfer editing activity approximately 2-fold, with little change in both the aminoacylation and the pretransfer editing activities. Ref.11
	Mutagenesis	394	1	D → A: Reduces the posttransfer editing activity approximately 2-fold, with little change in both the aminoacylation and the pretransfer editing activities. Ref.11
	Mutagenesis	443	1	C → G: 150-fold decrease in posttransfer editing activity against alanine, without change neither in pretransfer editing nor in aminoacylation. Ref.9
	Sequence conflict	26 – 27	2	ML → IV Ref.1
	Sequence conflict	127 – 128	2	QL → HV in AAA23710. Ref.2
	Sequence conflict	205 – 216	12	SASHE…VLAQS → RPLTNSRCWRR in AAA23710. Ref.2
	Sequence conflict	205	1	S → Q Ref.1
	Sequence conflict	516 – 517	2	PG → RA Ref.2
	Sequence conflict	518 – 572	55	Missing Ref.2

Sequences

	Sequence		Length	Mass (Da)
	P16659-1 [UniParc]. Last modified August 29, 2003. Version 4. Checksum: 312D29390E91C486	FASTA	572	63,693
	10 20 30 40 50 60 MRTSQYLLST LKETPADAEV ISHQLMLRAG MIRKLASGLY TWLPTGVRVL KKVENIVREE 70 80 90 100 110 120 MNNAGAIEVS MPVVQPADLW QESGRWEQYG PELLRFVDRG ERPFVLGPTH EEVITDLIRN 130 140 150 160 170 180 ELSSYKQLPL NFYQIQTKFR DEVRPRFGVM RSREFLMKDA YSFHTSQESL QETYDAMYAA 190 200 210 220 230 240 YSKIFSRMGL DFRAVQADTG SIGGSASHEF QVLAQSGEDD VVFSDTSDYA ANIELAEAIA 250 260 270 280 290 300 PKEPRAAATQ EMTLVDTPNA KTIAELVEQF NLPIEKTVKT LLVKAVEGSS FPQVALLVRG 310 320 330 340 350 360 DHELNEVKAE KLPQVASPLT FATEEEIRAV VKAGPGSLGP VNMPIPVVID RTVAAMSDFA 370 380 390 400 410 420 AGANIDGKHY FGINWDRDVA TPEVADIRNV VAGDPSPDGQ GRLLIKRGIE VGHIFQLGTK 430 440 450 460 470 480 YSEALKASVQ GEDGRNQILT MGCYGIGVTR VVAAAIEQNY DERGIVWPDA IAPFQVAILP 490 500 510 520 530 540 MNMHKSFRVQ ELAEKLYSEL RAQGIEVLLD DRKERPGVMF ADMELIGIPH TIVLGDRNLD 550 560 570 NDDIEYKYRR NGEKQLIKTG DIVEYLVKQI KG

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Partition of tRNA synthetases into two classes based on mutually exclusive sets of sequence motifs." Eriani G., Delarue M., Poch O., Gangloff J., Moras D. Nature 347:203-206(1990) [PubMed: 2203971] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-12.
[2]	"Identification and sequence of the drpA gene from Escherichia coli." Zhou Z., Syvanen M. J. Bacteriol. 172:281-286(1990) [PubMed: 1688424] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[3]	"Systematic sequencing of the Escherichia coli genome: analysis of the 4.0 - 6.0 min (189,987 - 281,416bp) region." Takemoto K., Mori H., Murayama N., Kataoka K., Yano M., Itoh T., Yamamoto Y., Inokuchi H., Miki T., Hatada E., Fukuda R., Ichihara S., Mizuno T., Makino K., Nakata A., Yura T., Sampei G., Mizobuchi K. Submitted (FEB-1996) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[4]	"Sequence of minutes 4-25 of Escherichia coli." Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M., Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D., Namath A., Oefner P., Roberts D., Schramm S., Davis R.W. Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[5]	"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.
[6]	"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], SEQUENCE REVISION TO 26-27; 205 AND 568-572. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[7]	Miyamoto K. Submitted (APR-1993) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-155. Strain: K12.
[8]	"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: 9298646] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-12. Strain: K12 / EMG2.
[9]	"Hydrolytic editing by a class II aminoacyl-tRNA synthetase." Beuning P.J., Musier-Forsyth K. Proc. Natl. Acad. Sci. U.S.A. 97:8916-8920(2000) [PubMed: 10922054] [Abstract] Cited for: EDITING ACTIVITY, KINETIC PARAMETERS, MUTAGENESIS OF CYS-443.
[10]	"Species-specific differences in amino acid editing by class II prolyl-tRNA synthetase." Beuning P.J., Musier-Forsyth K. J. Biol. Chem. 276:30779-30785(2001) [PubMed: 11408489] [Abstract] Cited for: EDITING ACTIVITY, KINETIC PARAMETERS.
[11]	"Functional role of the prokaryotic proline-tRNA synthetase insertion domain in amino acid editing." Wong F.-C., Beuning P.J., Nagan M., Shiba K., Musier-Forsyth K. Biochemistry 41:7108-7115(2002) [PubMed: 12033945] [Abstract] Cited for: MUTAGENESIS OF THR-257; LYS-279; ASP-350; HIS-369; ASP-378; ASP-386 AND ASP-394.
[12]	"Cysteine activation is an inherent in vitro property of prolyl-tRNA synthetases." Ahel I., Stathopoulos C., Ambrogelly A., Sauerwald A., Toogood H., Hartsch T., Soell D. J. Biol. Chem. 277:34743-34748(2002) [PubMed: 12130657] [Abstract] Cited for: PROLINE AND CYSTEINE ACTIVATION, EDITING ACTIVITY, KINETIC PARAMETERS.
[13]	"An isolated class II aminoacyl-tRNA synthetase insertion domain is functional in amino acid editing." Wong F.-C., Beuning P.J., Silvers C., Musier-Forsyth K. J. Biol. Chem. 278:52857-52864(2003) [PubMed: 14530268] [Abstract] Cited for: EDITING DOMAIN.

Cross-references

Sequence databases
EMBLGenBankDDBJ	X55518 Genomic DNA. Translation: CAA39134.1. M97858 Genomic DNA. Translation: AAA24420.1. M32357 Genomic DNA. Translation: AAA23710.1. U70214 Genomic DNA. Translation: AAB08622.1. U00096 Genomic DNA. Translation: AAC73305.1. AP009048 Genomic DNA. Translation: BAA77870.2. D15061 Genomic DNA. Translation: BAA03654.1.
PIR	YPEC. B64744.
RefSeq	AP_000855.1. NP_414736.1.
3D structure databases
ModBase	Search...
Protein-protein interaction databases
DIP	DIP:10573N.
IntAct	P16659. 14 interactions.
STRING	P16659.
2-D gel databases
SWISS-2DPAGE	P16659.
2DBase-Ecoli	P16659.
Proteomic databases
PRIDE	P16659.
Genome annotation databases
GeneID	949116.
GenomeReviews	Gene locus JW0190 in contig AP009048_GR. Gene locus b0194 in contig U00096_GR.
KEGG	ecj:JW0190. eco:b0194.
Organism-specific databases
EchoBASE	EB0763.
EcoGene	EG10770. proS.
CMR	Search...
Phylogenomic databases
HOGENOM	P16659.
OMA	VVSHQLM.
Enzyme and pathway databases
BioCyc	EcoCyc:PROS-MON. MetaCyc:PROS-MON.
Gene expression databases
Genevestigator	P16659.
Family and domain databases
HAMAP	MF_01569. [Tree]
InterPro	IPR002314. aa-tRNA-synt_IIb_cons-reg. IPR006195. aa-tRNA-synth_II_cons-reg. IPR004154. Anticodon_bd. IPR004500. Pro-tRNA-synth_IIa_bac. IPR002316. Pro-tRNA-synth_IIa_cons-reg. IPR007214. YbaK/aa-tRNA-synth-assoc-reg. [Graphical view]
Gene3D	G3DSA:3.40.50.800. Anticodon_bd. 1 hit.
Pfam	PF03129. HGTP_anticodon. 1 hit. PF00587. tRNA-synt_2b. 1 hit. PF04073. YbaK. 1 hit. [Graphical view]
PRINTS	PR01046. TRNASYNTHPRO.
TIGRFAMs	TIGR00409. proS_fam_II. 1 hit.
PROSITE	PS50862. AA_TRNA_LIGASE_II. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

SYP_ECOLI

Accession

Primary (citable) accession number: P16659
Secondary accession number(s): P78272, Q59430

Entry history

Integrated into UniProtKB/Swiss-Prot:	August 1, 1990
Last sequence update:	August 29, 2003
Last modified:	November 3, 2009
This is version 102 of the entry and version 4 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

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