Q93NG6 (DHPON_ARTNI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names	Recommended name: 2,6-dihydropseudooxynicotine hydrolase EC=3.7.1.19
Encoded on	Plasmid pAO1
Organism	Arthrobacter nicotinovorans
Taxonomic identifier	29320 [NCBI]
Taxonomic lineage	cellular organisms › Bacteria › Actinobacteria › Actinobacteria › Actinobacteridae › Actinomycetales › Micrococcineae › Micrococcaceae › Arthrobacter

Protein attributes

Sequence length	367 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	L-nicotine is used as a growth substrate. Plays a role in nicotine catabolism by cleaving a C-C bond in 2,6-dihydroxypseudooxynicotine. Ref.4 Ref.5
Catalytic activity	1-(2,6-dihydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O = 2,6-dihydroxypyridine + 4-methylaminobutanoate. Ref.4 Ref.5
Pathway	Alkaloid degradation; nicotine degradation; 2,6-dihydroxypyridine and 4-(methylamino)butanoate from 6-hydroxypseudooxynicotine: step 2/2. Ref.3 Ref.4 Ref.5
Subunit structure	Homodimer. Ref.5
Sequence similarities	Belongs to the UPF0255 family.
Biophysicochemical properties	Kinetic parameters: KM=6 µM for 2,6-dihydropseudooxynicotine Ref.4 pH dependence: Optimum pH is 7.5. Ref.4

Ontologies

Keywords
Biological process	Alkaloid metabolism
Molecular function	Hydrolase
Technical term	3D-structure Plasmid
Gene Ontology (GO)
Biological_process	nicotine catabolic process Inferred from electronic annotation. Source: UniProtKB-UniPathway
Molecular_function	hydrolase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 367

367

2,6-dihydropseudooxynicotine hydrolase

PRO_0000283735

Sites

Active site

148

1

Potential Ref.5

Active site

217

1

Ref.5

Active site

300

1

Ref.5

Active site

329

1

Ref.5

Experimental info

Mutagenesis

18

1

R → A: Total loss of activity. Ref.5

Mutagenesis

22

1

D → A: 91% loss of activity. Ref.5

Mutagenesis

148

1

E → A: 98% loss of activity. Ref.5

Mutagenesis

149

1

S → A: 84% loss of activity. Ref.5

Mutagenesis

153

1

E → A: 32% loss of activity. Ref.5

Mutagenesis

216

1

R → A: 92% loss of activity. Ref.5

Mutagenesis

217

1

S → A: Total loss of activity. Ref.5

Mutagenesis

300

1

D → A: 94% loss of activity. Ref.5

Mutagenesis

329

1

H → A: 92% loss of activity. Ref.5

Mutagenesis

330

1

C → A: 5% loss of activity. Ref.5

Mutagenesis

332

1

H → A: 58% loss of activity. Ref.5

Secondary structure

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367

Helix Strand Turn

Details...

Helix

9 – 13

5

Helix

16 – 22

7

Helix

26 – 33

8

Helix

41 – 61

21

Helix

65 – 83

19

Helix

90 – 105

16

Helix

106 – 108

3

Beta strand

109 – 111

3

Beta strand

113 – 120

8

Beta strand

123 – 130

8

Beta strand

133 – 135

3

Beta strand

138 – 144

7

Turn

151 – 153

3

Helix

155 – 163

9

Beta strand

167 – 171

5

Helix

177 – 179

3

Turn

180 – 182

3

Helix

189 – 202

14

Beta strand

206 – 216

11

Helix

218 – 229

12

Beta strand

235 – 240

6

Helix

248 – 250

3

Helix

253 – 262

10

Helix

268 – 278

11

Turn

282 – 284

3

Helix

285 – 287

3

Beta strand

292 – 297

6

Beta strand

300 – 302

3

Helix

305 – 313

9

Helix

316 – 318

3

Beta strand

319 – 324

6

Helix

329 – 334

6

Helix

337 – 350

14

Beta strand

360 – 362

3

Sequences

	Sequence		Length	Mass (Da)
	Q93NG6 [UniParc]. Last modified December 1, 2001. Version 1. Checksum: 1A5D31A7DFC4C8E3	FASTA	367	40,995
	10 20 30 40 50 60 MTVTSQVKPE DEMLNWGRLI LDGVSYSDMV GARDRPKEIT WFDYWMSLAN EYEQEAERKV 70 80 90 100 110 120 ALGHDLSAGE LLMSAALCAQ YAQFLWFDER RQKGQARKVE LYQKAAPLLS PPAERHELVV 130 140 150 160 170 180 DGIPMPVYVR IPEGPGPHPA VIMLGGLEST KEESFQMENL VLDRGMATAT FDGPGQGEMF 190 200 210 220 230 240 EYKRIAGDYE KYTSAVVDLL TKLEAIRNDA IGVLGRSLGG NYALKSAACE PRLAACISWG 250 260 270 280 290 300 GFSDLDYWDL ETPLTKESWK YVSKVDTLEE ARLHVHAALE TRDVLSQIAC PTYILHGVHD 310 320 330 340 350 360 EVPLSFVDTV LELVPAEHLN LVVEKDGDHC CHNLGIRPRL EMADWLYDVL VAGKKVAPTM KGWPLNG

References

[1]	"Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in degradation of the plant alkaloid nicotine: cloning, purification, and characterization of 2,6-dihydroxypyridine 3-hydroxylase." Baitsch D., Sandu C., Brandsch R., Igloi G.L. J. Bacteriol. 183:5262-5267(2001) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"Sequence of the 165-kilobase catabolic plasmid pAO1 from Arthrobacter nicotinovorans and identification of a pAO1-dependent nicotine uptake system." Igloi G.L., Brandsch R. J. Bacteriol. 185:1976-1986(2003) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[3]	"The bacterial oxidation of nicotine. VI. The metabolism of 2,6-dihydroxypseudooxynicotine." Gherna R.L., Richardson S.H., Rittenberg S.C. J. Biol. Chem. 240:3669-3674(1965) [PubMed] [Europe PMC] [Abstract] Cited for: PATHWAY.
[4]	"An alpha/beta-fold C--C bond hydrolase is involved in a central step of nicotine catabolism by Arthrobacter nicotinovorans." Sachelaru P., Schiltz E., Igloi G.L., Brandsch R. J. Bacteriol. 187:8516-8519(2005) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY.
[5]	"Structure and action of a C-C bond cleaving alpha/beta-hydrolase involved in nicotine degradation." Schleberger C., Sachelaru P., Brandsch R., Schulz G.E. J. Mol. Biol. 367:409-418(2007) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY, PATHWAY, SUBUNIT, ACTIVE SITE, MUTAGENESIS OF ARG-18; ASP-22; GLU-148; SER-149; GLU-153; ARG-216; SER-217; ASP-300; HIS-329; CYS-330 AND HIS-332.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AF373840 Genomic DNA. Translation: AAK64252.1.
AJ507836 Genomic DNA. Translation: CAD47941.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2JBW	X-ray	2.10	A/B/C/D	1-365	[»]

ProteinModelPortal

Q93NG6.

SMR

Q93NG6. Positions 15-365.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Enzyme and pathway databases

SABIO-RK

Q93NG6.

UniPathway

UPA00106; UER00490.

Family and domain databases

InterPro

IPR010520. DUF1100_hydro-like.
[Graphical view]

Pfam

PF06500. DUF1100. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

Q93NG6.

Entry information

Entry name

DHPON_ARTNI

Accession

Primary (citable) accession number: Q93NG6

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 3, 2007
Last sequence update:	December 1, 2001
Last modified:	May 1, 2013
This is version 50 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program