Q9I596 (NCASE_PSEAE) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Neutral ceramidase
Short name=N-CDase
Short name=NCDase
EC=3.5.1.23

Alternative name(s):
Acylsphingosine deacylase
N-acylsphingosine amidohydrolase

Gene names

Ordered Locus Names:

PA0845

Organism

Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)

Taxonomic identifier

208964 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Pseudomonadales › Pseudomonadaceae › Pseudomonas

Protein attributes

Sequence length	670 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	Hydrolyzes the ceramides into sphingosine and free fatty acid at an optimal pH of 8-9.
Catalytic activity	N-acylsphingosine + H2O = a carboxylate + sphingosine. Ref.1 Ref.3 Ref.4
Enzyme regulation	Inhibited by EDTA, EGTA and D/L-sphinganine D-erythro-sphingosine. L-erythro-sphingosine is a less powerful inhibitor. Stimulated by glycerophospholipids: cardiolipin is the most effective, followed by phosphatidic acid, phosphatidylethanolamine and phosphatidylglycerol, whereas phosphatidylcholine, lysophosphatidic acid and diacylglycerol are less effective. Ref.5
Subcellular location	Secreted Potential Ref.3.
Miscellaneous	Alternate N-termini have been proposed by different authors. It either starts from Asp-25 (Ref.1) or from Leu-27 (Ref.3).
Sequence similarities	Belongs to the neutral ceramidase family.
Biophysicochemical properties	Kinetic parameters: KM=139 µM for N-palmitoylsphingosine Ref.3 Ref.4 Vmax=5.3 µmol/min/mg enzyme with N-palmitoylsphingosine as substrate pH dependence: Optimum pH is 7.5-9.5.

Ontologies

Keywords
Biological process	Lipid metabolism
Cellular component	Secreted
Domain	Signal
Molecular function	Hydrolase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological process	lipid metabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	ceramidase 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

Signal peptide

1 – 24

24

Ref.1

Chain

25 – 670

646

Neutral ceramidase

PRO_0000247111

Regions

Region

644 – 670

27

Required for correct folding and localization

Sites

Active site

274

1

Nucleophile By similarity

Experimental info

Mutagenesis

665

1

V → D: Abolishes enzyme activity. Ref.6

Sequence conflict

90

1

I → T in BAA88409. Ref.1

Sequence conflict

181

1

N → S in BAA88409. Ref.1

Sequence conflict

196

1

V → A in BAA88409. Ref.1

Sequence conflict

598

1

E → V in BAA88409. Ref.1

Secondary structure

1

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670

Helix Strand Turn

Details...

Beta strand

28 – 31

4

Beta strand

65 – 74

10

Beta strand

79 – 85

7

Helix

92 – 105

14

Turn

112 – 114

3

Beta strand

115 – 119

5

Helix

133 – 136

4

Helix

137 – 139

3

Helix

144 – 151

8

Helix

155 – 162

8

Beta strand

170 – 172

3

Beta strand

179 – 184

6

Helix

186 – 189

4

Beta strand

207 – 212

6

Beta strand

222 – 224

3

Helix

247 – 253

7

Beta strand

264 – 266

3

Beta strand

273 – 275

3

Helix

289 – 295

7

Helix

299 – 307

9

Beta strand

313 – 315

3

Beta strand

320 – 327

8

Helix

335 – 337

3

Helix

374 – 378

5

Helix

389 – 394

6

Turn

395 – 397

3

Beta strand

408 – 410

3

Beta strand

415 – 423

9

Beta strand

427 – 430

4

Beta strand

434 – 436

3

Helix

443 – 452

10

Beta strand

459 – 462

4

Beta strand

466 – 468

3

Helix

476 – 481

6

Helix

495 – 512

18

Helix

526 – 528

3

Beta strand

550 – 552

3

Beta strand

556 – 559

4

Beta strand

563 – 569

7

Helix

573 – 575

3

Beta strand

583 – 588

6

Beta strand

598 – 602

5

Beta strand

608 – 615

8

Beta strand

619 – 627

9

Beta strand

637 – 641

5

Turn

648 – 650

3

Beta strand

663 – 666

4

Sequences

	Sequence		Length	Mass (Da)
	Q9I596 [UniParc]. Last modified March 1, 2001. Version 1. Checksum: F12C73EAC9CED287	FASTA	670	73,373
	10 20 30 40 50 60 MSRSAFTALL LSCVLLALSM PARADDLPYR FGLGKADITG EAAEVGMMGY SSLEQKTAGI 70 80 90 100 110 120 HMRQWARAFV IEEAASGRRL VYVNTDLGMI FQAVHLKVLA RLKAKYPGVY DENNVMLAAT 130 140 150 160 170 180 HTHSGPGGFS HYAMYNLSVL GFQEKTFNAI VDGIVRSIER AQARLQPGRL FYGSGELRNA 190 200 210 220 230 240 NRNRSLLSHL KNPDIVGYED GIDPQMSVLS FVDANGELAG AISWFPVHST SMTNANHLIS 250 260 270 280 290 300 PDNKGYASYH WEHDVSRKSG FVAAFAQTNA GNLSPNLNLK PGSGPFDNEF DNTREIGLRQ 310 320 330 340 350 360 FAKAYEIAGQ AQEEVLGELD SRFRFVDFTR LPIRPEFTDG QPRQLCTAAI GTSLAAGSTE 370 380 390 400 410 420 DGPGPLGLEE GNNPFLSALG GLLTGVPPQE LVQCQAEKTI LADTGNKKPY PWTPTVLPIQ 430 440 450 460 470 480 MFRIGQLELL GAPAEFTVMA GVRIRRAVQA ASEAAGIRHV VFNGYANAYA SYVTTREEYA 490 500 510 520 530 540 AQEYEGGSTL YGPWTQAAYQ QLFVDMAVAL RERLPVETSA IAPDLSCCQM NFQTGVVADD 550 560 570 580 590 600 PYIGKSFGDV LQQPRESYRI GDKVTVAFVT GHPKNDLRTE KTFLEVVNIG KDGKQTPETV 610 620 630 640 650 660 ATDNDWDTQY RWERVGISAS KATISWSIPP GTEPGHYYIR HYGNAKNFWT QKISEIGGST 670 RSFEVLGTTP

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Molecular cloning, sequencing, and expression of the gene encoding alkaline ceramidase from Pseudomonas aeruginosa. Cloning of a ceramidase homologue from Mycobacterium tuberculosis." Okino N., Ichinose S., Omori A., Imayama S., Nakamura T., Ito M. J. Biol. Chem. 274:36616-36622(1999) [PubMed: 10593963] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 25-48; 546-574; 583-591 AND 653-668, ENZYME ACTIVITY.
[2]	"Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen." Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P., Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M., Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y., Brody L.L., Coulter S.N., Folger K.R. , Kas A., Larbig K., Lim R.M., Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J., Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V. Nature 406:959-964(2000) [PubMed: 10984043] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228.
[3]	"Molecular cloning and characterization of the alkaline ceramidase from Pseudomonas aeruginosa PA01." Nieuwenhuizen W.F., van Leeuwen S., Jack R.W., Egmond M.R., Goetz F. Protein Expr. Purif. 30:94-104(2003) [PubMed: 12821326] [Abstract] Cited for: PROTEIN SEQUENCE OF 26-58, ENZYME ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR LOCATION. Strain: ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228.
[4]	"Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa." Okino N., Tani M., Imayama S., Ito M. J. Biol. Chem. 273:14368-14373(1998) [PubMed: 9603946] [Abstract] Cited for: ENZYME ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES.
[5]	"Activation of bacterial ceramidase by anionic glycerophospholipids: possible involvement in ceramide hydrolysis on atopic skin by Pseudomonas ceramidase." Kita K., Sueyoshi N., Okino N., Inagaki M., Ishida H., Kiso M., Imayama S., Nakamura T., Ito M. Biochem. J. 362:619-626(2002) [PubMed: 11879188] [Abstract] Cited for: ENZYME REGULATION.
[6]	"Conserved amino acid residues in the COOH-terminal tail are indispensable for the correct folding and localization and enzyme activity of neutral ceramidase." Tani M., Okino N., Sueyoshi N., Ito M. J. Biol. Chem. 279:29351-29358(2004) [PubMed: 15123644] [Abstract] Cited for: MUTAGENESIS OF VAL-665.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AB028646 Genomic DNA. Translation: BAA88409.1.
AJ315932 Genomic DNA. Translation: CAC67511.1.
AE004091 Genomic DNA. Translation: AAG04234.1.

PIR

C83540.

RefSeq

NP_249536.1. NC_002516.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2ZWS	X-ray	1.40	A	25-670	[»]
2ZXC	X-ray	2.20	A/B	25-670	[»]

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

GeneID

880698.

GenomeReviews

Gene locus PA0845 in contig AE004091_GR.

KEGG

pae:PA0845.

NMPDR

fig|208964.1.peg.846.

PATRIC

19835970. VBIPseAer58763_0877.

Organism-specific databases

PseudoCAP

PA0845.

Phylogenomic databases

HOGENOM

HBG315824.

OMA

SWFAVHP.

ProtClustDB

CLSK866161.

Enzyme and pathway databases

BioCyc

PAER208964:PA0845-MONOMER.

BRENDA

3.5.1.23. 5087.

Family and domain databases

InterPro

IPR006823. Ceramidase_alk.
[Graphical view]

PANTHER

PTHR12670. Ceramidase_alk. 1 hit.

Pfam

PF04734. Ceramidase_alk. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

NCASE_PSEAE

Accession

Primary (citable) accession number: Q9I596
Secondary accession number(s): Q7AY51, Q9RHQ0

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 25, 2006
Last sequence update:	March 1, 2001
Last modified:	January 25, 2012
This is version 59 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program