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Q91XT9 (ASAH2_RAT) Reviewed, UniProtKB/Swiss-Prot

Last modified November 16, 2011. Version 60. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (1) | Third-party data text xml rdf/xml gff fasta
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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 2
N-acylsphingosine amidohydrolase 2

Cleaved into the following chain:

  1. Neutral ceramidase soluble form
Gene names
Name:Asah2
OrganismRattus norvegicus (Rat)
Taxonomic identifier10116 [NCBI]
Taxonomic lineageEukaryota › Metazoa › Chordata › Craniata › Vertebrata › Euteleostomi › Mammalia › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Rattus

Protein attributes

Sequence length761 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Hydrolyzes the sphingolipid ceramide into sphingosine and free fatty acid at an optimal pH of 6.5-8.5. Acts as a key regulator of sphingolipid signaling metabolites by generating sphingosine at the cell surface. Acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine, a precursor of the antiapoptotic factor sphingosine 1-phosphate. Probably involved in the digestion of dietary sphingolipids in intestine by acting as a key enzyme for the catabolism of dietary sphingolipids and regulating the levels of bioactive sphingolipid metabolites in the intestinal tract. Ref.10

Catalytic activity

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

Enzyme regulation

Inhibited by dithiothreitol (DTT), 2-mercaptoethanol, Zn2+ and Cu2+.

Subcellular location

Cell membrane; Single-pass type II membrane protein. Note: The neutral ceramidase soluble form is a secreted protein. Ref.1 Ref.2

Tissue specificity

Widelky expressed. Highly expressed in brain, kidney and heart. Expressed at lower level in other tissues such as liver. Localizes in the epithelia of the jejunum and ileum. Ref.1 Ref.5

Induction

By interleukin-1-beta in renal mesengial cells. Ref.6

Post-translational modification

N-glycosylated. Required for enzyme activity. Ref.1 Ref.2 Ref.10

O-glycosylated. Required to retain it as a type II membrane protein at the cell surface. Ref.1 Ref.2 Ref.10

Phosphorylated. May prevent ubiquitination and subsequent degradation. Ref.7

Ubiquitinated, leading to its degradation by the proteasome. Ubiquitination is triggered by nitric oxid. Ref.8

Sequence similarities

Belongs to the neutral ceramidase family.

Biophysicochemical properties

Kinetic parameters:

KM=71.4 µM for octanoyl-sphingosine Ref.1 Ref.4 Ref.10

KM=66 µM for palmitoyl-sphingosine

KM=1.29 M for C16-ceramide

KM=3.84 M for dihydroceramide

Vmax=160 µmol/min/mg enzyme with octanoyl-sphingosine as substrate

Vmax=16 µmol/min/mg enzyme with palmitoyl-sphingosine as substrate

Vmax=4.4 µmol/min/mg enzyme with C16-ceramide as substrate

Vmax=1.2 µmol/min/mg enzyme with dihydroceramide as substrate

pH dependence:

Optimum pH is 7-10.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 761761Neutral ceramidase
PRO_0000247103
Chain80 – 761682Neutral ceramidase soluble form
PRO_0000247104

Regions

Topological domain1 – 1111Cytoplasmic Potential
Transmembrane12 – 3221Helical; Signal-anchor for type II membrane protein; Potential
Topological domain33 – 761729Lumenal Potential
Region751 – 76111Required for correct folding and localization

Sites

Active site3351Nucleophile By similarity

Amino acid modifications

Glycosylation511O-linked (GalNAc...) Potential
Glycosylation521O-linked (GalNAc...) Potential
Glycosylation561O-linked (GalNAc...) Potential
Glycosylation571O-linked (GalNAc...) Potential
Glycosylation581O-linked (GalNAc...) Potential
Glycosylation601O-linked (GalNAc...) Potential
Glycosylation611O-linked (GalNAc...) Potential
Glycosylation631O-linked (GalNAc...) Potential
Glycosylation641O-linked (GalNAc...) Potential
Glycosylation661O-linked (GalNAc...) Potential
Glycosylation681O-linked (GalNAc...) Potential
Glycosylation691O-linked (GalNAc...) Potential
Glycosylation711O-linked (GalNAc...) Potential
Glycosylation1981N-linked (GlcNAc...) Potential
Glycosylation4121N-linked (GlcNAc...) Potential
Glycosylation4491N-linked (GlcNAc...) Potential

Experimental info

Mutagenesis441S → A: Abolishes O-glycosylation and localization at the cell surface; when associated with A-50; A-51; A-52; A-56; A-57; A-58; A-60; A-61; A-62; A-63; A-65; A-76; A-68; A-69; A-71 and A-77. Ref.2
Mutagenesis50 – 7122STTQG…PTTQT → AAAQGPAAAQAAPAAQAPAA QA: Abolishes O-glycosylation and localization at the cell surface; when associated with A-44 and A-77. Ref.2
Mutagenesis771S → A: Abolishes O-glycosylation and localization at the cell surface; when associated with A-44; A-50; A-51; A-52; A-56; A-57; A-58; A-60; A-61; A-62; A-63; A-65; A-76; A-68; A-69 and A-71. Ref.2
Mutagenesis7561F → I: No effect. Ref.9
Mutagenesis7561F → R or D: Loss of function. Ref.9
Mutagenesis7571E → R: No effect. Ref.9
Mutagenesis7581I → F: Impairs enzyme activity. Ref.9
Mutagenesis7581I → R or D: Loss of function. Ref.9
Mutagenesis7581I → V: No effect. Ref.9
Sequence conflict3251G → N AA sequence Ref.3

Sequences

Sequence LengthMass (Da)Tools
Q91XT9 [UniParc].

Last modified December 1, 2001. Version 1.
Checksum: 68B91BC78AEB6324

FASTA76183,488
        10         20         30         40         50         60 
MAKRTFSSLE AFLIFLLVMM TAITVALLTL LFVTSGTIEN HKDSGNHWVS TTQGPTTTQS 

        70         80         90        100        110        120 
SPTTQTPTTQ TPDLPPSQNF SGYYIGVGRA DCTGQVSDIN LMGYGKNGQN AQGLLTRLFS 

       130        140        150        160        170        180 
RAFILADPDG SNRMAFVSVE LCMISQRLRL EVLKRLQSKY GSLYRRDNVI LSATHTHSGP 

       190        200        210        220        230        240 
AGFFQYTLYI LASEGFSNRT FQYIVSGIVK SIDIAHTNLK PGKVLINKGN VANVQINRSP 

       250        260        270        280        290        300 
SSYLQNPPSE RARYSSDTDK EMVVLKLVDL NGEDLGLISW FAVHPVSMNN SNHLVNSDNM 

       310        320        330        340        350        360 
GYAAYLFEQE KNRGYLPGQG PFVAGFASSN LGDVSPNILG PHCVNTGESC DNDKSTCPSG 

       370        380        390        400        410        420 
GPSMCMASGP GQDMFESTHI IGRVIYQKAK ELHASASQEV TGPVLTAHQW VNMTDVSVQL 

       430        440        450        460        470        480 
NATHTVKTCK AALGYSFAAG TIDGVSGLNI TQGTTEGNLF WDTLRDQLLG KPSEEIIECQ 

       490        500        510        520        530        540 
KPKPILIHTG ELTKPHPWQP DIVDIQIVTL GSLAIAAIPG EFTTMSGRRL REAVKKEFAL 

       550        560        570        580        590        600 
YGMKDMTVVI AGLSNVYTHY ITTYEEYQAQ RYEAASTIYG PHTLSAYIQL FRALAKAIAT 

       610        620        630        640        650        660 
DTVANMSSGP EPPFFKNLIG SLIPNIADRA PIGKQFGDVL QPAKPEYRVG EVVEVVFVGA 

       670        680        690        700        710        720 
NPKNSAENQT HQTFLTVEKY EDSVANWQIM HNDASWETRF YWHKGVLGLS NATIHWHIPD 

       730        740        750        760 
TALPGVYRIR YFGHNRKQEL LKPAVILAFE GISSPFEIVT T

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References

[1]"Purification, characterization, molecular cloning, and subcellular distribution of neutral ceramidase of rat kidney."
Mitsutake S., Tani M., Okino N., Mori K., Ichinose S., Omori A., Iida H., Nakamura T., Ito M.
J. Biol. Chem. 276:26249-26259(2001) [PubMed: 11328816] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 223-245; 261-273; 601-619 AND 701-729, ENZYME ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, GLYCOSYLATION, SUBCELLULAR LOCATION, TISSUE SPECIFICITY.
Tissue: Kidney.
[2]"O-glycosylation of mucin-like domain retains the neutral ceramidase on the plasma membranes as a type II integral membrane protein."
Tani M., Iida H., Ito M.
J. Biol. Chem. 278:10523-10530(2003) [PubMed: 12499379] [Abstract]
Cited for: PROTEIN SEQUENCE OF 80-100, SUBCELLULAR LOCATION, TOPOLOGY, GLYCOSYLATION, MUTAGENESIS OF SER-44; 50-SER--THR-71 AND SER-77.
[3]"Molecular cloning and characterization of a human mitochondrial ceramidase."
El Bawab S., Roddy P., Qian T., Bielawska A., Lemasters J.J., Hannun Y.A.
J. Biol. Chem. 275:21508-21513(2000) [PubMed: 10781606] [Abstract]
Cited for: PROTEIN SEQUENCE OF 311-327; 635-648 AND 737-753.
[4]"Purification and characterization of a membrane-bound nonlysosomal ceramidase from rat brain."
El Bawab S., Bielawska A., Hannun Y.A.
J. Biol. Chem. 274:27948-27955(1999) [PubMed: 10488143] [Abstract]
Cited for: ENZYME ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES.
[5]"Distribution and properties of neutral ceramidase activity in rat intestinal tract."
Lundgren P., Nilsson A., Duan R.D.
Dig. Dis. Sci. 46:765-772(2001) [PubMed: 11330410] [Abstract]
Cited for: TISSUE SPECIFICITY.
[6]"Interleukin-1beta induces chronic activation and de novo synthesis of neutral ceramidase in renal mesangial cells."
Franzen R., Pautz A., Braeutigam L., Geisslinger G., Pfeilschifter J., Huwiler A.
J. Biol. Chem. 276:35382-35389(2001) [PubMed: 11457826] [Abstract]
Cited for: INDUCTION.
[7]"Nitric oxide induces degradation of the neutral ceramidase in rat renal mesangial cells and is counterregulated by protein kinase C."
Franzen R., Fabbro D., Aschrafi A., Pfeilschifter J., Huwiler A.
J. Biol. Chem. 277:46184-46190(2002) [PubMed: 12359735] [Abstract]
Cited for: PHOSPHORYLATION.
[8]"Nitric oxide induces neutral ceramidase degradation by the ubiquitin/proteasome complex in renal mesangial cell cultures."
Franzen R., Pfeilschifter J., Huwiler A.
FEBS Lett. 532:441-444(2002) [PubMed: 12482609] [Abstract]
Cited for: UBIQUITINATION.
[9]"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 PHE-756; GLU-757 AND ILE-758.
[10]"Rat intestinal ceramidase: purification, properties, and physiological relevance."
Olsson M., Duan R.-D., Ohlsson L., Nilsson A.
Am. J. Physiol. 287:G929-G937(2004) [PubMed: 15217782] [Abstract]
Cited for: IDENTIFICATION BY MASS PSECTROMETRY, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, GLYCOSYLATION.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ		AB057433 mRNA. Translation: BAB62033.1.
IPIIPI00208165.
RefSeqNP_446098.1. NM_053646.1.
UniGeneRn.156958.

3D structure databases

ModBaseSearch...

Protein-protein interaction databases

STRINGQ91XT9.

Proteomic databases

PRIDEQ91XT9.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

GeneID114104.
KEGGrno:114104.

Organism-specific databases

CTD56624.
RGD69410. Asah2.

Phylogenomic databases

eggNOGroNOG12354.
GeneTreeENSGT00390000015792.
HOVERGENHBG080870.
InParanoidQ91XT9.
OrthoDBEOG4G4GPW.

Enzyme and pathway databases

BRENDA3.5.1.23. 5301.

Gene expression databases

ArrayExpressQ91XT9.
GenevestigatorQ91XT9.
GermOnlineENSRNOG00000012196. Rattus norvegicus.

Family and domain databases

InterProIPR006823. Ceramidase_alk.
[Graphical view]
KOK12349.
PANTHERPTHR12670. Ceramidase_alk. 1 hit.
PfamPF04734. Ceramidase_alk. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

NextBio618273.

Entry information

Entry nameASAH2_RAT
AccessionPrimary (citable) accession number: Q91XT9
Entry history
Integrated into UniProtKB/Swiss-Prot: July 25, 2006
Last sequence update: December 1, 2001
Last modified: November 16, 2011
This is version 60 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programChordata Protein Annotation Program

Relevant documents

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