P77044 (MHPC_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
2-hydroxy-6-oxononadienedioate/2-hydroxy-6-oxononatrienedioate hydrolase
EC=3.7.1.14

Alternative name(s):
2-hydroxy-6-ketonona-2,4-diene-1,9-dioic acid 5,6-hydrolase
2-hydroxy-6-oxonona-2,4,7-triene-1,9-dioic acid 5,6-hydrolase
2-hydroxy-6-oxonona-2,4-diene-1,9-dioic acid 5,6-hydrolase

Gene names

Name:	mhpC
Ordered Locus Names:	b0349, JW0340

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	288 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	Catalyzes the cleavage of the C5-C6 bond of 2-hydroxy-6-oxononadienedioate and 2-hydroxy-6-oxononatrienedioate, a dienol ring fission product of the bacterial meta-cleavage pathway for degradation of phenylpropionic acid. MhpC shows some selectivity for the carboxylate of the side chain. Ref.2 Ref.6
Catalytic activity	(2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + succinate. HAMAP-Rule MF_01654 (2Z,4E,7E)-2-hydroxy-6-oxonona-2,4,7-triene-1,9-dioate + H2O = (2Z)-2-hydroxypenta-2,4-dienoate + fumarate. HAMAP-Rule MF_01654
Pathway	Aromatic compound metabolism; 3-phenylpropanoate degradation. HAMAP-Rule MF_01654
Subunit structure	Homodimer. Ref.6 Ref.9
Miscellaneous	MhpC is not a serine hydrolase (catalytic triad), as Ser-114 is a non-nucleophilic catalytic residue and Asp-239 is not involved in the catalytic mechanism. HAMAP-Rule MF_01654
Sequence similarities	Belongs to the AB hydrolase superfamily. MhpC family.
Biophysicochemical properties	Kinetic parameters: KM=2.1 µM for 2-hydroxy-6-oxononadienedioate (at pH 8) Ref.6
Mass spectrometry	Molecular mass is 31717 Da from positions 8 - 288. Determined by ESI. Ref.9
Sequence caution	The sequence AAB18073.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened. The sequence AAC73452.2 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened. The sequence BAA13054.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened. The sequence BAE76131.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened. The sequence CAA70749.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally shortened.

Ontologies

Keywords
Biological process	Aromatic hydrocarbons catabolism
Molecular function	Hydrolase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	3-(3-hydroxy)phenylpropionate catabolic process Inferred from mutant phenotype PubMed 3531186. Source: EcoCyc 3-phenylpropionate catabolic process Inferred from electronic annotation. Source: UniProtKB-UniPathway aromatic compound catabolic process Inferred from mutant phenotype PubMed 3531186. Source: EcoliWiki
Cellular_component	cytoplasm Inferred from direct assay Ref.6. Source: EcoliWiki
Molecular_function	2-hydroxy-6-oxonona-2,4,7-trienedioate hydrolase activity Inferred from direct assay Ref.6. Source: EcoCyc 2-hydroxy-6-oxonona-2,4-dienedioate hydrolase activity Inferred from direct assay Ref.6. Source: EcoCyc
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed Ref.6

Chain

2 – 288

287

2-hydroxy-6-oxononadienedioate/2-hydroxy-6-oxononatrienedioate hydrolase HAMAP-Rule MF_01654

PRO_0000207054

Sites

Active site

267

1

Proton acceptor

Site

114

1

Transition state stabilizer

Site

192

1

Catalytic role in ketonization of the dienol substrate (substrate destabilization)

Experimental info

Mutagenesis

44

1

S → A: 2-fold decrease in catalytic efficiency and more than 5-fold increase in affinity for the natural substrate. Ref.7

Mutagenesis

113

1

N → A: 200-fold decrease in catalytic activity and almost 2-fold increase in affinity. Ref.8

Mutagenesis

113

1

N → H: 350-fold decrease in catalytic activity and almost 2-fold increase in affinity. Ref.8

Mutagenesis

114

1

S → A: Weakly active. 3-fold decrease in affinity. Fast ketonisation and slow C-C cleavage. Ref.7

Mutagenesis

114

1

S → G: Weakly active. 3-fold decrease in affinity. Fast ketonisation and slow C-C cleavage. Ref.7

Mutagenesis

118

1

H → A: More than 2-fold decrease in catalytic efficiency and 3-fold increase affinity. Ref.7

Mutagenesis

177

1

F → D: 100-fold decrease in catalytic activity. Ref.8

Mutagenesis

177

1

F → G: 4-fold and 8-fold decrease in catalytic activity and affinity, respectively. Ref.8

Mutagenesis

192

1

R → K: 40-fold and 5-fold decrease in catalytic activity and affinity, respectively. Ref.8

Mutagenesis

192

1

R → Q: 280-fold and 10-fold decrease in catalytic activity and affinity, respectively. Ref.8

Mutagenesis

265

1

C → A: 2-fold decrease in catalytic activity and almost 2-fold increase in affinity. Ref.8

Mutagenesis

267

1

H → A: Weakly active, 1000-fold decrease in catalytic efficiency. Very slow ketonisation and C-C cleavage. Ref.7

Mutagenesis

268

1

W → G: 10-fold and 20-fold decrease in catalytic activity and affinity, respectively. Ref.8

Sequence conflict

153

1

E → G in BAA13054. Ref.1

Secondary structure

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288

Helix Strand Turn

Details...

Helix

8 – 11

4

Beta strand

12 – 19

8

Beta strand

22 – 31

10

Beta strand

35 – 41

7

Helix

50 – 53

4

Turn

54 – 57

4

Helix

58 – 63

6

Beta strand

67 – 71

5

Helix

88 – 102

15

Beta strand

108 – 113

6

Helix

115 – 126

12

Helix

128 – 130

3

Beta strand

131 – 138

8

Beta strand

146 – 148

3

Helix

153 – 163

11

Helix

167 – 175

9

Helix

186 – 198

13

Helix

200 – 212

13

Helix

221 – 226

6

Beta strand

231 – 236

6

Beta strand

240 – 242

3

Helix

245 – 253

9

Beta strand

258 – 264

7

Helix

269 – 272

4

Helix

274 – 285

12

Sequences

	Sequence		Length	Mass (Da)
	P77044 [UniParc]. Last modified December 14, 2011. Version 4. Checksum: 8F7E5ADE584A45BA	FASTA	288	31,937
	10 20 30 40 50 60 MSYQPQTEAA TSRFLNVEEA GKTLRIHFND CGQGDETVVL LHGSGPGATG WANFSRNIDP 70 80 90 100 110 120 LVEAGYRVIL LDCPGWGKSD SVVNSGSRSD LNARILKSVV DQLDIAKIHL LGNSMGGHSS 130 140 150 160 170 180 VAFTLKWPER VGKLVLMGGG TGGMSLFTPM PTEGIKRLNQ LYRQPTIENL KLMMDIFVFD 190 200 210 220 230 240 TSDLTDALFE ARLNNMLSRR DHLENFVKSL EANPKQFPDF GPRLAEIKAQ TLIVWGRNDR 250 260 270 280 FVPMDAGLRL LSGIAGSELH IFRDCGHWAQ WEHADAFNQL VLNFLARP

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	Kawamukai M. Submitted (JUN-1996) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[2]	"Genetic characterization and expression in heterologous hosts of the 3-(3-hydroxyphenyl)propionate catabolic pathway of Escherichia coli K-12." Ferrandez A., Garcia J.L., Diaz E. J. Bacteriol. 179:2573-2581(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION AS A HYDROLASE. Strain: K12 / CS520.
[3]	"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.
[4]	"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.
[5]	"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.
[6]	"Purification, characterization, and stereochemical analysis of a C-C hydrolase: 2-hydroxy-6-keto-nona-2,4-diene-1,9-dioic acid 5,6-hydrolase." Lam W.W., Bugg T.D. Biochemistry 36:12242-12251(1997) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-18, FUNCTION AS A HYDROLASE, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.
[7]	"Catalytic mechanism of C-C hydrolase MhpC from Escherichia coli: kinetic analysis of His263 and Ser110 site-directed mutants." Li C., Montgomery M.G., Mohammed F., Li J.-J., Wood S.P., Bugg T.D.H. J. Mol. Biol. 346:241-251(2005) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF SER-44; SER-114; HIS-118 AND HIS-267, CATALYTIC MECHANISM.
[8]	"Catalytic role for arginine 188 in the C-C hydrolase catalytic mechanism for Escherichia coli MhpC and Burkholderia xenovorans LB400 BphD." Li C., Li J.-J., Montgomery M.G., Wood S.P., Bugg T.D.H. Biochemistry 45:12470-12479(2006) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF ASN-113; PHE-177; ARG-192; CYS-265 AND TRP-268, CATALYTIC MECHANISM.
[9]	"The structure of the C-C bond hydrolase MhpC provides insights into its catalytic mechanism." Dunn G., Montgomery M.G., Mohammed F., Coker A., Cooper J.B., Robertson T., Garcia J.-L., Bugg T.D.H., Wood S.P. J. Mol. Biol. 346:253-265(2005) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 5-288 IN COMPLEX WITH A NON-CLEAVABLE SUBSTRATE ANALOG, MASS SPECTROMETRY, SUBUNIT.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

D86239 Genomic DNA. Translation: BAA13054.1. Different initiation.
Y09555 Genomic DNA. Translation: CAA70749.1. Different initiation.
U73857 Genomic DNA. Translation: AAB18073.1. Different initiation.
U00096 Genomic DNA. Translation: AAC73452.2. Different initiation.
AP009048 Genomic DNA. Translation: BAE76131.1. Different initiation.

PIR

E64762.

RefSeq

NP_414883.5. NC_000913.2.
YP_488643.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1U2E	X-ray	2.10	A/B/C/D	1-288	[»]

ProteinModelPortal

P77044.

SMR

P77044. Positions 3-288.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-10207N.

IntAct

P77044. 3 interactions.

STRING

511145.b0349.

Protein family/group databases

MEROPS

S33.995.

Proteomic databases

PaxDb

P77044.

PRIDE

P77044.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC73452; AAC73452; b0349.
BAE76131; BAE76131; BAE76131.

GeneID

12931735.
944954.

KEGG

ecj:Y75_p0338.
eco:b0349.

PATRIC

32115831. VBIEscCol129921_0357.

Organism-specific databases

EchoBASE

EB4168.

EcoGene

EG20275. mhpC.

Phylogenomic databases

eggNOG

COG0596.

HOGENOM

HOG000028063.

KO

K05714.

ProtClustDB

CLSK879654.

Enzyme and pathway databases

BioCyc

EcoCyc:MHPCHYDROL-MONOMER.
ECOL316407:JW0340-MONOMER.
MetaCyc:MHPCHYDROL-MONOMER.

UniPathway

UPA00714.

Gene expression databases

Genevestigator

P77044.

Family and domain databases

HAMAP

MF_01654. MhpC.

InterPro

IPR000073. AB_hydrolase_1.
IPR000639. Epox_hydrolase-like.
IPR023791. MhpC_alpha/beta_hydrolase.
[Graphical view]

PRINTS

PR00111. ABHYDROLASE.
PR00412. EPOXHYDRLASE.

ProtoNet

Search...

Other

EvolutionaryTrace

P77044.

Entry information

Entry name

MHPC_ECOLI

Accession

Primary (citable) accession number: P77044
Secondary accession number(s): P71204, P77205, Q2MC75

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1997
Last sequence update:	December 14, 2011
Last modified:	May 1, 2013
This is version 111 of the entry and version 4 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program