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Protein

Neutral ceramidase

Gene

Asah2

Organism
Rattus norvegicus (Rat)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

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.1 Publication

Catalytic activityi

N-acylsphingosine + H2O = a carboxylate + sphingosine.2 Publications

Enzyme regulationi

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

Kineticsi

  1. KM=71.4 µM for octanoyl-sphingosine3 Publications
  2. KM=66 µM for palmitoyl-sphingosine3 Publications
  3. KM=1.29 M for C16-ceramide3 Publications
  4. KM=3.84 M for dihydroceramide3 Publications
  1. Vmax=160 µmol/min/mg enzyme with octanoyl-sphingosine as substrate3 Publications
  2. Vmax=16 µmol/min/mg enzyme with palmitoyl-sphingosine as substrate3 Publications
  3. Vmax=4.4 µmol/min/mg enzyme with C16-ceramide as substrate3 Publications
  4. Vmax=1.2 µmol/min/mg enzyme with dihydroceramide as substrate3 Publications

pH dependencei

Optimum pH is 7-10.3 Publications

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Active sitei335 – 3351NucleophileBy similarity

GO - Molecular functioni

GO - Biological processi

  • apoptotic process Source: UniProtKB-KW
  • response to organic substance Source: RGD
  • sphingolipid metabolic process Source: UniProtKB-KW
Complete GO annotation...

Keywords - Molecular functioni

Hydrolase

Keywords - Biological processi

Apoptosis, Lipid metabolism, Sphingolipid metabolism

Enzyme and pathway databases

BRENDAi3.5.1.23. 5301.
ReactomeiR-RNO-1660662. Glycosphingolipid metabolism.
SABIO-RKQ91XT9.

Chemistry

SwissLipidsiSLP:000000682.

Names & Taxonomyi

Protein namesi
Recommended name:
Neutral ceramidase (EC:3.5.1.23)
Short name:
N-CDase
Short name:
NCDase
Alternative name(s):
Acylsphingosine deacylase 2
N-acylsphingosine amidohydrolase 2
Cleaved into the following chain:
Gene namesi
Name:Asah2
OrganismiRattus norvegicus (Rat)
Taxonomic identifieri10116 [NCBI]
Taxonomic lineageiEukaryotaMetazoaChordataCraniataVertebrataEuteleostomiMammaliaEutheriaEuarchontogliresGliresRodentiaSciurognathiMuroideaMuridaeMurinaeRattus
Proteomesi
  • UP000002494 Componenti: Chromosome 1

Organism-specific databases

RGDi69410. Asah2.

Subcellular locationi

Topology

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Topological domaini1 – 1111CytoplasmicSequence analysisAdd
BLAST
Transmembranei12 – 3221Helical; Signal-anchor for type II membrane proteinSequence analysisAdd
BLAST
Topological domaini33 – 761729LumenalSequence analysisAdd
BLAST

GO - Cellular componenti

Complete GO annotation...

Keywords - Cellular componenti

Cell membrane, Membrane

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi44 – 441S → 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. 1 Publication
Mutagenesisi50 – 7122STTQG…PTTQT → AAAQGPAAAQAAPAAQAPAA QA: Abolishes O-glycosylation and localization at the cell surface; when associated with A-44 and A-77. 1 PublicationAdd
BLAST
Mutagenesisi77 – 771S → 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. 1 Publication
Mutagenesisi756 – 7561F → I: No effect. 1 Publication
Mutagenesisi756 – 7561F → R or D: Loss of function. 1 Publication
Mutagenesisi757 – 7571E → R: No effect. 1 Publication
Mutagenesisi758 – 7581I → F: Impairs enzyme activity. 1 Publication
Mutagenesisi758 – 7581I → R or D: Loss of function. 1 Publication
Mutagenesisi758 – 7581I → V: No effect. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 761761Neutral ceramidasePRO_0000247103Add
BLAST
Chaini80 – 761682Neutral ceramidase soluble formPRO_0000247104Add
BLAST

Amino acid modifications

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Glycosylationi51 – 511O-linked (GalNAc...)Sequence analysis
Glycosylationi52 – 521O-linked (GalNAc...)Sequence analysis
Glycosylationi56 – 561O-linked (GalNAc...)Sequence analysis
Glycosylationi57 – 571O-linked (GalNAc...)Sequence analysis
Glycosylationi58 – 581O-linked (GalNAc...)Sequence analysis
Glycosylationi60 – 601O-linked (GalNAc...)Sequence analysis
Glycosylationi61 – 611O-linked (GalNAc...)Sequence analysis
Glycosylationi63 – 631O-linked (GalNAc...)Sequence analysis
Glycosylationi64 – 641O-linked (GalNAc...)Sequence analysis
Glycosylationi66 – 661O-linked (GalNAc...)Sequence analysis
Glycosylationi68 – 681O-linked (GalNAc...)Sequence analysis
Glycosylationi69 – 691O-linked (GalNAc...)Sequence analysis
Glycosylationi71 – 711O-linked (GalNAc...)Sequence analysis
Glycosylationi198 – 1981N-linked (GlcNAc...)Sequence analysis
Glycosylationi412 – 4121N-linked (GlcNAc...)Sequence analysis
Glycosylationi449 – 4491N-linked (GlcNAc...)Sequence analysis

Post-translational modificationi

N-glycosylated. Required for enzyme activity.
O-glycosylated. Required to retain it as a type II membrane protein at the cell surface.
Phosphorylated. May prevent ubiquitination and subsequent degradation.2 Publications
Ubiquitinated, leading to its degradation by the proteasome. Ubiquitination is triggered by nitric oxid.1 Publication

Keywords - PTMi

Glycoprotein, Phosphoprotein, Ubl conjugation

Proteomic databases

PaxDbiQ91XT9.
PRIDEiQ91XT9.

Expressioni

Tissue specificityi

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.2 Publications

Inductioni

By interleukin-1-beta in renal mesengial cells.1 Publication

Gene expression databases

GenevisibleiQ91XT9. RN.

Interactioni

Protein-protein interaction databases

STRINGi10116.ENSRNOP00000016688.

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni751 – 76111Required for correct folding and localizationAdd
BLAST

Sequence similaritiesi

Belongs to the neutral ceramidase family.Curated

Keywords - Domaini

Signal-anchor, Transmembrane, Transmembrane helix

Phylogenomic databases

eggNOGiKOG2232. Eukaryota.
ENOG410XQWE. LUCA.
GeneTreeiENSGT00390000015792.
HOGENOMiHOG000209915.
HOVERGENiHBG080870.
InParanoidiQ91XT9.
KOiK12349.
OMAiMEDWFEN.
OrthoDBiEOG7WQ7RQ.
PhylomeDBiQ91XT9.

Family and domain databases

InterProiIPR006823. Ceramidase_alk.
IPR031331. NEUT/ALK_ceramidase_C.
IPR031329. NEUT/ALK_ceramidase_N.
[Graphical view]
PANTHERiPTHR12670. PTHR12670. 2 hits.
PfamiPF04734. Ceramidase_alk. 1 hit.
PF17048. Ceramidse_alk_C. 1 hit.
[Graphical view]

Sequencei

Sequence statusi: Complete.

Sequence processingi: The displayed sequence is further processed into a mature form.

Q91XT9-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MAKRTFSSLE AFLIFLLVMM TAITVALLTL LFVTSGTIEN HKDSGNHWVS
60 70 80 90 100
TTQGPTTTQS SPTTQTPTTQ TPDLPPSQNF SGYYIGVGRA DCTGQVSDIN
110 120 130 140 150
LMGYGKNGQN AQGLLTRLFS RAFILADPDG SNRMAFVSVE LCMISQRLRL
160 170 180 190 200
EVLKRLQSKY GSLYRRDNVI LSATHTHSGP AGFFQYTLYI LASEGFSNRT
210 220 230 240 250
FQYIVSGIVK SIDIAHTNLK PGKVLINKGN VANVQINRSP SSYLQNPPSE
260 270 280 290 300
RARYSSDTDK EMVVLKLVDL NGEDLGLISW FAVHPVSMNN SNHLVNSDNM
310 320 330 340 350
GYAAYLFEQE KNRGYLPGQG PFVAGFASSN LGDVSPNILG PHCVNTGESC
360 370 380 390 400
DNDKSTCPSG GPSMCMASGP GQDMFESTHI IGRVIYQKAK ELHASASQEV
410 420 430 440 450
TGPVLTAHQW VNMTDVSVQL NATHTVKTCK AALGYSFAAG TIDGVSGLNI
460 470 480 490 500
TQGTTEGNLF WDTLRDQLLG KPSEEIIECQ KPKPILIHTG ELTKPHPWQP
510 520 530 540 550
DIVDIQIVTL GSLAIAAIPG EFTTMSGRRL REAVKKEFAL YGMKDMTVVI
560 570 580 590 600
AGLSNVYTHY ITTYEEYQAQ RYEAASTIYG PHTLSAYIQL FRALAKAIAT
610 620 630 640 650
DTVANMSSGP EPPFFKNLIG SLIPNIADRA PIGKQFGDVL QPAKPEYRVG
660 670 680 690 700
EVVEVVFVGA NPKNSAENQT HQTFLTVEKY EDSVANWQIM HNDASWETRF
710 720 730 740 750
YWHKGVLGLS NATIHWHIPD TALPGVYRIR YFGHNRKQEL LKPAVILAFE
760
GISSPFEIVT T
Length:761
Mass (Da):83,488
Last modified:December 1, 2001 - v1
Checksum:i68B91BC78AEB6324
GO

Experimental Info

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Sequence conflicti325 – 3251G → N AA sequence (PubMed:10781606).Curated

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AB057433 mRNA. Translation: BAB62033.1.
RefSeqiNP_446098.1. NM_053646.2.
XP_006231323.1. XM_006231261.2.
XP_006231325.1. XM_006231263.1.
XP_006231329.1. XM_006231267.2.
XP_006231330.1. XM_006231268.2.
XP_006231332.1. XM_006231270.1.
XP_008758541.1. XM_008760319.1.
XP_008758542.1. XM_008760320.1.
UniGeneiRn.156958.

Genome annotation databases

EnsembliENSRNOT00000016688; ENSRNOP00000016688; ENSRNOG00000012196.
ENSRNOT00000077135; ENSRNOP00000073116; ENSRNOG00000012196.
GeneIDi114104.
KEGGirno:114104.
UCSCiRGD:69410. rat.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AB057433 mRNA. Translation: BAB62033.1.
RefSeqiNP_446098.1. NM_053646.2.
XP_006231323.1. XM_006231261.2.
XP_006231325.1. XM_006231263.1.
XP_006231329.1. XM_006231267.2.
XP_006231330.1. XM_006231268.2.
XP_006231332.1. XM_006231270.1.
XP_008758541.1. XM_008760319.1.
XP_008758542.1. XM_008760320.1.
UniGeneiRn.156958.

3D structure databases

ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

STRINGi10116.ENSRNOP00000016688.

Chemistry

SwissLipidsiSLP:000000682.

Proteomic databases

PaxDbiQ91XT9.
PRIDEiQ91XT9.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

EnsembliENSRNOT00000016688; ENSRNOP00000016688; ENSRNOG00000012196.
ENSRNOT00000077135; ENSRNOP00000073116; ENSRNOG00000012196.
GeneIDi114104.
KEGGirno:114104.
UCSCiRGD:69410. rat.

Organism-specific databases

CTDi56624.
RGDi69410. Asah2.

Phylogenomic databases

eggNOGiKOG2232. Eukaryota.
ENOG410XQWE. LUCA.
GeneTreeiENSGT00390000015792.
HOGENOMiHOG000209915.
HOVERGENiHBG080870.
InParanoidiQ91XT9.
KOiK12349.
OMAiMEDWFEN.
OrthoDBiEOG7WQ7RQ.
PhylomeDBiQ91XT9.

Enzyme and pathway databases

BRENDAi3.5.1.23. 5301.
ReactomeiR-RNO-1660662. Glycosphingolipid metabolism.
SABIO-RKQ91XT9.

Miscellaneous databases

NextBioi618273.
PROiQ91XT9.

Gene expression databases

GenevisibleiQ91XT9. RN.

Family and domain databases

InterProiIPR006823. Ceramidase_alk.
IPR031331. NEUT/ALK_ceramidase_C.
IPR031329. NEUT/ALK_ceramidase_N.
[Graphical view]
PANTHERiPTHR12670. PTHR12670. 2 hits.
PfamiPF04734. Ceramidase_alk. 1 hit.
PF17048. Ceramidse_alk_C. 1 hit.
[Graphical view]
ProtoNetiSearch...

Publicationsi

  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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [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] [Europe PMC] [Abstract]
    Cited for: IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, GLYCOSYLATION.

Entry informationi

Entry nameiASAH2_RAT
AccessioniPrimary (citable) accession number: Q91XT9
Entry historyi
Integrated into UniProtKB/Swiss-Prot: July 25, 2006
Last sequence update: December 1, 2001
Last modified: February 17, 2016
This is version 87 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programChordata Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

Complete proteome, Direct protein sequencing, Reference proteome

Documents

  1. SIMILARITY comments
    Index of protein domains and families

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.