NCASE_MYCTU
ID NCASE_MYCTU Reviewed; 637 AA.
AC O06769; F2GNL0; L0T7D5;
DT 28-NOV-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-1997, sequence version 1.
DT 03-AUG-2022, entry version 112.
DE RecName: Full=Neutral ceramidase;
DE Short=N-CDase;
DE Short=NCDase;
DE EC=3.5.1.23 {ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604};
DE AltName: Full=Acylsphingosine deacylase;
DE AltName: Full=N-acylsphingosine amidohydrolase;
GN OrderedLocusNames=Rv0669c;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP SEQUENCE REVISION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=12368430; DOI=10.1099/00221287-148-10-2967;
RA Camus J.-C., Pryor M.J., Medigue C., Cole S.T.;
RT "Re-annotation of the genome sequence of Mycobacterium tuberculosis
RT H37Rv.";
RL Microbiology 148:2967-2973(2002).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=10593963; DOI=10.1074/jbc.274.51.36616;
RA Okino N., Ichinose S., Omori A., Imayama S., Nakamura T., Ito M.;
RT "Molecular cloning, sequencing, and expression of the gene encoding
RT alkaline ceramidase from Pseudomonas aeruginosa. Cloning of a ceramidase
RT homologue from Mycobacterium tuberculosis.";
RL J. Biol. Chem. 274:36616-36622(1999).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, AND SUBSTRATE SPECIFICITY.
RX PubMed=20139604; DOI=10.1271/bbb.90645;
RA Okino N., Ikeda R., Ito M.;
RT "Expression, purification, and characterization of a recombinant neutral
RT ceramidase from Mycobacterium tuberculosis.";
RL Biosci. Biotechnol. Biochem. 74:316-321(2010).
RN [5]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
CC -!- FUNCTION: Catalyzes the cleavage of the N-acyl linkage of the ceramides
CC (Cers) to yield sphingosine (Sph) and free fatty acid. Also catalyzes
CC the synthesis of Cers from Sph and fatty acid. Cers containning C6-C24
CC fatty acids are well hydrolyzed, and Cers with mono unsaturated fatty
CC acids are much more hydrolyzed than those with saturated fatty acids.
CC {ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-acylsphing-4-enine + H2O = a fatty acid + sphing-4-enine;
CC Xref=Rhea:RHEA:20856, ChEBI:CHEBI:15377, ChEBI:CHEBI:28868,
CC ChEBI:CHEBI:52639, ChEBI:CHEBI:57756; EC=3.5.1.23;
CC Evidence={ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-acylsphinganine + H2O = a fatty acid + sphinganine;
CC Xref=Rhea:RHEA:33551, ChEBI:CHEBI:15377, ChEBI:CHEBI:28868,
CC ChEBI:CHEBI:31488, ChEBI:CHEBI:57817;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-acyl-(4R)-4-hydroxysphinganine + H2O = (4R)-
CC hydroxysphinganine + a fatty acid; Xref=Rhea:RHEA:33555,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:28868, ChEBI:CHEBI:31998,
CC ChEBI:CHEBI:64124; Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-(9Z-octadecenoyl)-sphing-4-enine = (9Z)-octadecenoate
CC + sphing-4-enine; Xref=Rhea:RHEA:41299, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:30823, ChEBI:CHEBI:57756, ChEBI:CHEBI:77996;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-(hexanoyl)sphing-4-enine = hexanoate + sphing-4-enine;
CC Xref=Rhea:RHEA:41295, ChEBI:CHEBI:15377, ChEBI:CHEBI:17120,
CC ChEBI:CHEBI:57756, ChEBI:CHEBI:63867;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-hexadecanoylsphing-4-enine = hexadecanoate + sphing-4-
CC enine; Xref=Rhea:RHEA:38891, ChEBI:CHEBI:7896, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:57756, ChEBI:CHEBI:72959;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-octadecanoylsphing-4-enine = octadecanoate + sphing-4-
CC enine; Xref=Rhea:RHEA:41279, ChEBI:CHEBI:15377, ChEBI:CHEBI:25629,
CC ChEBI:CHEBI:57756, ChEBI:CHEBI:72961;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-eicosanoyl-sphing-4-enine = eicosanoate + sphing-4-
CC enine; Xref=Rhea:RHEA:41275, ChEBI:CHEBI:15377, ChEBI:CHEBI:32360,
CC ChEBI:CHEBI:57756, ChEBI:CHEBI:72962;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-(15Z-tetracosenoyl)-sphing-4-enine = (15Z)-
CC tetracosenoate + sphing-4-enine; Xref=Rhea:RHEA:41267,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:32392, ChEBI:CHEBI:57756,
CC ChEBI:CHEBI:74450; Evidence={ECO:0000269|PubMed:20139604};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N-tetracosanoyl-sphing-4-enine = sphing-4-enine +
CC tetracosanoate; Xref=Rhea:RHEA:41283, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:31014, ChEBI:CHEBI:57756, ChEBI:CHEBI:72965;
CC Evidence={ECO:0000269|PubMed:20139604};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000250|UniProtKB:Q9I596};
CC Note=Binds 1 zinc ion per subunit. {ECO:0000250|UniProtKB:Q9I596};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q9I596};
CC -!- ACTIVITY REGULATION: 90% of activity is inhibited by nickel, zinc and
CC calcium ions. Magnesium, cobalt, copper and manganese ions inhibit
CC between 50 and 80% of activity. {ECO:0000269|PubMed:20139604}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=98.7 uM for N-dodecanoyl-7-nitrobenz-2-oxa-1,3-4-diazole (NBD)-D-
CC erythro-sphingosine (C12-NBD-Cer) (at pH 8 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604};
CC Vmax=21.1 pmol/min/mg enzyme (at pH 8 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604};
CC pH dependence:
CC Optimum pH is between 8 and 9. {ECO:0000269|PubMed:10593963,
CC ECO:0000269|PubMed:20139604};
CC Temperature dependence:
CC 10 minutes at 100 degrees Celsius abolishes completely the activity.
CC {ECO:0000269|PubMed:10593963, ECO:0000269|PubMed:20139604};
CC -!- SIMILARITY: Belongs to the neutral ceramidase family. {ECO:0000305}.
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DR EMBL; AL123456; CCP43412.1; -; Genomic_DNA.
DR PIR; H70535; H70535.
DR RefSeq; NP_215183.1; NC_000962.3.
DR RefSeq; WP_003900995.1; NZ_NVQJ01000007.1.
DR AlphaFoldDB; O06769; -.
DR SMR; O06769; -.
DR STRING; 83332.Rv0669c; -.
DR SwissLipids; SLP:000001155; -.
DR PaxDb; O06769; -.
DR DNASU; 888181; -.
DR GeneID; 888181; -.
DR KEGG; mtu:Rv0669c; -.
DR TubercuList; Rv0669c; -.
DR eggNOG; ENOG502Z84X; Bacteria.
DR InParanoid; O06769; -.
DR OMA; VWHRTNT; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005576; C:extracellular region; IBA:GO_Central.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; HDA:MTBBASE.
DR GO; GO:0102121; F:ceramidase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0017040; F:N-acylsphingosine amidohydrolase activity; IDA:MTBBASE.
DR GO; GO:0046514; P:ceramide catabolic process; IDA:MTBBASE.
DR GO; GO:0042759; P:long-chain fatty acid biosynthetic process; IDA:MTBBASE.
DR GO; GO:0046512; P:sphingosine biosynthetic process; IDA:MTBBASE.
DR Gene3D; 2.60.40.2300; -; 1.
DR InterPro; IPR006823; Ceramidase_alk.
DR InterPro; IPR038445; NCDase_C_sf.
DR InterPro; IPR031331; NEUT/ALK_ceramidase_C.
DR InterPro; IPR031329; NEUT/ALK_ceramidase_N.
DR PANTHER; PTHR12670; PTHR12670; 1.
DR Pfam; PF04734; Ceramidase_alk; 1.
DR Pfam; PF17048; Ceramidse_alk_C; 1.
PE 1: Evidence at protein level;
KW Hydrolase; Lipid metabolism; Magnesium; Metal-binding; Reference proteome;
KW Zinc.
FT CHAIN 1..637
FT /note="Neutral ceramidase"
FT /id="PRO_0000420250"
FT ACT_SITE 256
FT /note="Nucleophile"
FT /evidence="ECO:0000250"
FT BINDING 34
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 96
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 204
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 417
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 453
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 575
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 577
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
FT BINDING 580
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9I596"
SQ SEQUENCE 637 AA; 69490 MW; 5E2D915AB1E4FDB7 CRC64;
MLSVGRGIAD ITGEAADCGM LGYGKSDQRT AGIHQRLRSR AFVFRDDSQD GDARLLLIVA
ELPLPMQNVN EEVLRRLADL YGDTYSEQNT LITATHTHAG PGGYCGYLLY NLTTSGFRPA
TFAAIVDGIV ESVEHAHADV APAEVSLSHG ELYGASINRS PSAFDRNPPA DKAFFPKRVD
PHTTLVRIDR GEATVGVIHF FATHGTSMTN RNHLISGDNK GFAAYHWERT VGGADYLAGQ
PDFIAAFAQT NPGDMSPNVD GPLSPEAPPD REFDNTRRTG LCQFEDAFTQ LSGATPIGAG
IDARFTYVDL GSVLVRGEYT PDGEERRTGR PMFGAGAMAG TDEGPGFHGF RQGRNPFWDR
LSRAMYRLAR PTAAAQAPKG IVMPARLPNR IHPFVQEIVP VQLVRIGRLY LIGIPGEPTI
VAGLRLRRMV ASIVGADLAD VLCVGYTNAY IHYVTTPEEY LEQRYEGGST LFGRWELCAL
MQTVAELAEA MRDGRPVTLG RRPRPTRELS WVRGAPADAG SFGAVIAEPS ATYRPGQAVE
AVFVSALPNN DLRRGGTYLE VVRREGASWV RIADDGDWAT SFRWQRQGRA GSHVSIRWDV
PGDTTPGQYR IVHHGTARDR NGMLTAFSAT TREFTVV
