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Q9I596 (NCASE_PSEAE) Reviewed, UniProtKB/Swiss-Prot

Last modified May 1, 2013. Version 70. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended 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
OrganismPseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) [Reference proteome] [HAMAP]
Taxonomic identifier208964 [NCBI]
Taxonomic lineageBacteria › Proteobacteria › Gammaproteobacteria › Pseudomonadales › Pseudomonadaceae › Pseudomonas ›

Protein attributes

Sequence length670 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Catalyzes the cleavage of the N-acyl linkage of the ceramides (Cers) to yield sphingosine (Sph) and free fatty acid at an optimal pH of 8-9. Also catalyzes the synthesis of Cers from Sph and fatty acid. Ref.4 Ref.7

Catalytic activity

N-acylsphingosine + H2O = a carboxylate + sphingosine. Ref.1 Ref.3 Ref.4

Cofactor

Binds 1 zinc ion per subunit. Ref.4 Ref.7

Magnesium. Ref.4 Ref.7

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

Subunit structure

Homodimer. Ref.7

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 processLipid metabolism
   Cellular componentSecreted
   DomainSignal
   LigandMagnesium
Metal-binding
Zinc
   Molecular functionHydrolase
   PTMDisulfide bond
   Technical term3D-structure
Complete proteome
Direct protein sequencing
Reference proteome
Gene Ontology (GO)
   Biological_processlong-chain fatty acid biosynthetic process

Inferred from direct assay Ref.4. Source: UniProtKB

sphingosine biosynthetic process

Inferred from direct assay Ref.4. Source: UniProtKB

   Cellular_componentextracellular region

Inferred from electronic annotation. Source: UniProtKB-SubCell

   Molecular_functionceramidase activity

Inferred from direct assay Ref.4. Source: UniProtKB

metal ion binding

Inferred from electronic annotation. Source: UniProtKB-KW

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Signal peptide1 – 2424 Ref.1
Chain25 – 670646Neutral ceramidase
PRO_0000247111

Regions

Region644 – 67027Required for correct folding and localization

Sites

Active site2741Nucleophile By similarity
Metal binding611Magnesium; via carbonyl oxygen
Metal binding1211Zinc; via tele nitrogen
Metal binding2281Zinc; via tele nitrogen
Metal binding4351Zinc
Metal binding4721Zinc
Metal binding6031Magnesium
Metal binding6051Magnesium; via carbonyl oxygen
Metal binding6081Magnesium
Binding site841Substrate
Binding site921Substrate
Binding site1111Substrate
Binding site1301Substrate
Binding site4691Substrate
Binding site4721Substrate
Binding site4841Substrate
Binding site5351Substrate; via amide nitrogen

Amino acid modifications

Disulfide bond394 ↔ 528

Experimental info

Mutagenesis1211H → A: No ceramidase activity; when associated with A-123. Ref.7
Mutagenesis1231H → A: No ceramidase activity; when associated with A-121. Ref.7
Mutagenesis1841R → A: No ceramidase activity; when associated. Ref.7
Mutagenesis4351E → A: Slight ceramidase activity. Ref.7
Mutagenesis4721Y → A: No ceramidase activity; when associated. Ref.7
Mutagenesis4841Y → A: Slight ceramidase activity. Ref.7
Mutagenesis6651V → D: Abolishes enzyme activity. Ref.6
Sequence conflict901I → T in BAA88409. Ref.1
Sequence conflict1811N → S in BAA88409. Ref.1
Sequence conflict1961V → A in BAA88409. Ref.1
Sequence conflict5981E → V in BAA88409. Ref.1

Secondary structure

1 .................................................................................................................. 670
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q9I596 [UniParc].

Last modified March 1, 2001. Version 1.
Checksum: F12C73EAC9CED287

FASTA67073,373
        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

« Hide

References

« Hide '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] [Europe PMC] [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. expand/collapse author list , 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] [Europe PMC] [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] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE OF 26-58, CATALYTIC 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] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, COFACTOR, 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] [Europe PMC] [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] [Europe PMC] [Abstract]
Cited for: MUTAGENESIS OF VAL-665.
[7]"Mechanistic insights into the hydrolysis and synthesis of ceramide by neutral ceramidase."
Inoue T., Okino N., Kakuta Y., Hijikata A., Okano H., Goda H.M., Tani M., Sueyoshi N., Kambayashi K., Matsumura H., Kai Y., Ito M.
J. Biol. Chem. 284:9566-9577(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS) OF 25-670 COMPLEX WITH SUBSTRATE ANALOGS AND DIVALENT CATIONS, FUNCTION, MUTAGENESIS OF HIS-121; HIS-123; ARG-184; GLU-435; TYR-472 AND TYR-484, REACTION MECHANISM, COFACTOR, SUBUNIT.

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.
PIRC83540.
RefSeqNP_249536.1. NC_002516.2.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
2ZWSX-ray1.40A25-670[»]
2ZXCX-ray2.20A/B25-670[»]
ProteinModelPortalQ9I596.
SMRQ9I596. Positions 26-668.
ModBaseSearch...

Protein-protein interaction databases

STRING208964.PA0845.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

GeneID880698.
KEGGpae:PA0845.
PATRIC19835970. VBIPseAer58763_0877.

Organism-specific databases

PseudoCAPPA0845.

Phylogenomic databases

eggNOGNOG75118.
HOGENOMHOG000209915.
OMASWFAVHP.
ProtClustDBCLSK866161.

Enzyme and pathway databases

BRENDA3.5.1.23. 5087.

Family and domain databases

InterProIPR006823. Ceramidase_alk.
[Graphical view]
PANTHERPTHR12670. PTHR12670. 1 hit.
PfamPF04734. Ceramidase_alk. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceQ9I596.

Entry information

Entry nameNCASE_PSEAE
AccessionPrimary (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: May 1, 2013
This is version 70 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

SIMILARITY comments

Index of protein domains and families

to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·Documents