NIT2_HUMAN
ID NIT2_HUMAN Reviewed; 276 AA.
AC Q9NQR4; B2R9A3; D3DN47; Q8WUF0;
DT 26-FEB-2008, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2000, sequence version 1.
DT 03-AUG-2022, entry version 155.
DE RecName: Full=Omega-amidase NIT2;
DE EC=3.5.1.3 {ECO:0000269|PubMed:19595734, ECO:0000269|PubMed:22674578};
DE AltName: Full=Nitrilase homolog 2;
GN Name=NIT2; ORFNames=CUA002;
OS Homo sapiens (Human).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae;
OC Homo.
OX NCBI_TaxID=9606;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=10959838; DOI=10.1016/s0960-9822(00)00621-7;
RA Pace H.C., Hodawadekar S.C., Draganescu A., Huang J., Bieganowski P.,
RA Pekarsky Y., Croce C.M., Brenner C.;
RT "Crystal structure of the worm NitFhit Rosetta stone protein reveals a Nit
RT tetramer binding two Fhit dimers.";
RL Curr. Biol. 10:907-917(2000).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC TISSUE=Adrenal tumor;
RA Xu X., Yang Y., Gao G., Xiao H., Chen Z., Han Z.;
RL Submitted (MAY-2000) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RX PubMed=14702039; DOI=10.1038/ng1285;
RA Ota T., Suzuki Y., Nishikawa T., Otsuki T., Sugiyama T., Irie R.,
RA Wakamatsu A., Hayashi K., Sato H., Nagai K., Kimura K., Makita H.,
RA Sekine M., Obayashi M., Nishi T., Shibahara T., Tanaka T., Ishii S.,
RA Yamamoto J., Saito K., Kawai Y., Isono Y., Nakamura Y., Nagahari K.,
RA Murakami K., Yasuda T., Iwayanagi T., Wagatsuma M., Shiratori A., Sudo H.,
RA Hosoiri T., Kaku Y., Kodaira H., Kondo H., Sugawara M., Takahashi M.,
RA Kanda K., Yokoi T., Furuya T., Kikkawa E., Omura Y., Abe K., Kamihara K.,
RA Katsuta N., Sato K., Tanikawa M., Yamazaki M., Ninomiya K., Ishibashi T.,
RA Yamashita H., Murakawa K., Fujimori K., Tanai H., Kimata M., Watanabe M.,
RA Hiraoka S., Chiba Y., Ishida S., Ono Y., Takiguchi S., Watanabe S.,
RA Yosida M., Hotuta T., Kusano J., Kanehori K., Takahashi-Fujii A., Hara H.,
RA Tanase T.-O., Nomura Y., Togiya S., Komai F., Hara R., Takeuchi K.,
RA Arita M., Imose N., Musashino K., Yuuki H., Oshima A., Sasaki N.,
RA Aotsuka S., Yoshikawa Y., Matsunawa H., Ichihara T., Shiohata N., Sano S.,
RA Moriya S., Momiyama H., Satoh N., Takami S., Terashima Y., Suzuki O.,
RA Nakagawa S., Senoh A., Mizoguchi H., Goto Y., Shimizu F., Wakebe H.,
RA Hishigaki H., Watanabe T., Sugiyama A., Takemoto M., Kawakami B.,
RA Yamazaki M., Watanabe K., Kumagai A., Itakura S., Fukuzumi Y., Fujimori Y.,
RA Komiyama M., Tashiro H., Tanigami A., Fujiwara T., Ono T., Yamada K.,
RA Fujii Y., Ozaki K., Hirao M., Ohmori Y., Kawabata A., Hikiji T.,
RA Kobatake N., Inagaki H., Ikema Y., Okamoto S., Okitani R., Kawakami T.,
RA Noguchi S., Itoh T., Shigeta K., Senba T., Matsumura K., Nakajima Y.,
RA Mizuno T., Morinaga M., Sasaki M., Togashi T., Oyama M., Hata H.,
RA Watanabe M., Komatsu T., Mizushima-Sugano J., Satoh T., Shirai Y.,
RA Takahashi Y., Nakagawa K., Okumura K., Nagase T., Nomura N., Kikuchi H.,
RA Masuho Y., Yamashita R., Nakai K., Yada T., Nakamura Y., Ohara O.,
RA Isogai T., Sugano S.;
RT "Complete sequencing and characterization of 21,243 full-length human
RT cDNAs.";
RL Nat. Genet. 36:40-45(2004).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RA Mural R.J., Istrail S., Sutton G.G., Florea L., Halpern A.L., Mobarry C.M.,
RA Lippert R., Walenz B., Shatkay H., Dew I., Miller J.R., Flanigan M.J.,
RA Edwards N.J., Bolanos R., Fasulo D., Halldorsson B.V., Hannenhalli S.,
RA Turner R., Yooseph S., Lu F., Nusskern D.R., Shue B.C., Zheng X.H.,
RA Zhong F., Delcher A.L., Huson D.H., Kravitz S.A., Mouchard L., Reinert K.,
RA Remington K.A., Clark A.G., Waterman M.S., Eichler E.E., Adams M.D.,
RA Hunkapiller M.W., Myers E.W., Venter J.C.;
RL Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA], AND VARIANT ALA-231.
RC TISSUE=Testis;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [6]
RP TISSUE SPECIFICITY, AND IDENTIFICATION BY MASS SPECTROMETRY.
RX PubMed=14528910;
RA Myung J.K., Gulesserian T., Fountoulakis M., Lubec G.;
RT "Deranged hypothetical proteins Rik protein, Nit protein 2 and
RT mitochondrial inner membrane protein, Mitofilin, in fetal Down syndrome
RT brain.";
RL Cell. Mol. Biol. 49:739-746(2003).
RN [7]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=15592455; DOI=10.1038/nbt1046;
RA Rush J., Moritz A., Lee K.A., Guo A., Goss V.L., Spek E.J., Zhang H.,
RA Zha X.-M., Polakiewicz R.D., Comb M.J.;
RT "Immunoaffinity profiling of tyrosine phosphorylation in cancer cells.";
RL Nat. Biotechnol. 23:94-101(2005).
RN [8]
RP TISSUE SPECIFICITY, SUBCELLULAR LOCATION, AND MASS SPECTROMETRY.
RX PubMed=17488281; DOI=10.1111/j.1742-4658.2007.05828.x;
RA Lin C.-H., Chung M.-Y., Chen W.-B., Chien C.-H.;
RT "Growth inhibitory effect of the human NIT2 gene and its allelic imbalance
RT in cancers.";
RL FEBS J. 274:2946-2956(2007).
RN [9]
RP FUNCTION, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, AND CATALYTIC ACTIVITY.
RX PubMed=19595734; DOI=10.1016/j.biochi.2009.07.003;
RA Krasnikov B.F., Chien C.-H., Nostramo R., Pinto J.T., Nieves E.,
RA Callaway M., Sun J., Huebner K., Cooper A.J.L.;
RT "Identification of the putative tumor suppressor Nit2 as omega-amidase, an
RT enzyme metabolically linked to glutamine and asparagine transamination.";
RL Biochimie 91:1072-1080(2009).
RN [10]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=21269460; DOI=10.1186/1752-0509-5-17;
RA Burkard T.R., Planyavsky M., Kaupe I., Breitwieser F.P., Buerckstuemmer T.,
RA Bennett K.L., Superti-Furga G., Colinge J.;
RT "Initial characterization of the human central proteome.";
RL BMC Syst. Biol. 5:17-17(2011).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, IDENTIFICATION
RP BY MASS SPECTROMETRY, ACTIVE SITE, AND MUTAGENESIS OF GLU-43; LYS-112;
RP CYS-153 AND 116-PHE--GLU-128.
RX PubMed=22674578; DOI=10.1074/jbc.m111.259119;
RA Chien C.H., Gao Q.Z., Cooper A.J., Lyu J.H., Sheu S.Y.;
RT "Structural insights into the catalytic active site and activity of human
RT Nit2/omega-amidase: kinetic assay and molecular dynamics simulation.";
RL J. Biol. Chem. 287:25715-25726(2012).
RN [12]
RP PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-26, AND IDENTIFICATION BY
RP MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC TISSUE=Liver;
RX PubMed=24275569; DOI=10.1016/j.jprot.2013.11.014;
RA Bian Y., Song C., Cheng K., Dong M., Wang F., Huang J., Sun D., Wang L.,
RA Ye M., Zou H.;
RT "An enzyme assisted RP-RPLC approach for in-depth analysis of human liver
RT phosphoproteome.";
RL J. Proteomics 96:253-262(2014).
RN [13]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=25944712; DOI=10.1002/pmic.201400617;
RA Vaca Jacome A.S., Rabilloud T., Schaeffer-Reiss C., Rompais M., Ayoub D.,
RA Lane L., Bairoch A., Van Dorsselaer A., Carapito C.;
RT "N-terminome analysis of the human mitochondrial proteome.";
RL Proteomics 15:2519-2524(2015).
CC -!- FUNCTION: Has omega-amidase activity (PubMed:22674578,
CC PubMed:19595734). The role of omega-amidase is to remove potentially
CC toxic intermediates by converting 2-oxoglutaramate and 2-oxosuccinamate
CC to biologically useful 2-oxoglutarate and oxaloacetate, respectively
CC (PubMed:19595734). {ECO:0000269|PubMed:19595734,
CC ECO:0000269|PubMed:22674578}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a monoamide of a dicarboxylate + H2O = a dicarboxylate +
CC NH4(+); Xref=Rhea:RHEA:11716, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:28965, ChEBI:CHEBI:77450; EC=3.5.1.3;
CC Evidence={ECO:0000269|PubMed:19595734, ECO:0000269|PubMed:22674578};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:11717;
CC Evidence={ECO:0000269|PubMed:19595734, ECO:0000269|PubMed:22674578};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutaramate + H2O = 2-oxoglutarate + NH4(+);
CC Xref=Rhea:RHEA:32963, ChEBI:CHEBI:15377, ChEBI:CHEBI:16769,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:28938; EC=3.5.1.3;
CC Evidence={ECO:0000269|PubMed:19595734, ECO:0000269|PubMed:22674578};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:32964;
CC Evidence={ECO:0000269|PubMed:19595734, ECO:0000269|PubMed:22674578};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxosuccinamate + H2O = NH4(+) + oxaloacetate;
CC Xref=Rhea:RHEA:59412, ChEBI:CHEBI:15377, ChEBI:CHEBI:16452,
CC ChEBI:CHEBI:28938, ChEBI:CHEBI:57735; EC=3.5.1.3;
CC Evidence={ECO:0000269|PubMed:22674578};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:59413;
CC Evidence={ECO:0000269|PubMed:22674578};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.009 mM for 2-oxoglutaramate (open-chain form at pH 8.5 and 37
CC degrees Celsius) {ECO:0000269|PubMed:19595734};
CC KM=0.003 mM for 2-oxoglutaramate (open-chain form at pH 8.5 and 37
CC degrees Celsius) {ECO:0000269|PubMed:22674578};
CC KM=10.9 mM for succinamate (at pH 7.2 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:22674578};
CC Vmax=5.9 umol/min/mg enzyme with 2-oxoglutaramate as substrate (open-
CC chain form at pH 8.5 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:19595734};
CC Vmax=30.3 umol/min/mg enzyme with 2-oxoglutaramate as substrate
CC (open-chain form at pH 8.5 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:22674578};
CC Vmax=169 umol/min/mg enzyme with succinamate as substrate (at pH 7.2
CC and 37 degrees Celsius) {ECO:0000269|PubMed:22674578};
CC Note=In solution, 2-oxoglutaramate is in equilibrium with a cyclic
CC form (2-hydroxy-5-oxoproline), and at pH 8.0 or above, the rate of
CC ring opening is no longer limiting for the omega-amidase reaction
CC (PubMed:19595734, PubMed:22674578). The kinetic constants are
CC determined for the recombinant His6-tagged protein (PubMed:22674578).
CC {ECO:0000303|PubMed:19595734, ECO:0000303|PubMed:22674578};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:19595734}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:17488281}.
CC -!- TISSUE SPECIFICITY: Detected in fetal brain (at protein level).
CC Ubiquitous. Detected in heart, brain, placenta, lung, liver, skeletal
CC muscle, kidney, pancreas, prostate, spleen, thymus, prostate, testis,
CC ovary, small intestine and colon. {ECO:0000269|PubMed:14528910,
CC ECO:0000269|PubMed:17488281}.
CC -!- MASS SPECTROMETRY: Mass=30585; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:17488281};
CC -!- SIMILARITY: Belongs to the carbon-nitrogen hydrolase superfamily.
CC NIT1/NIT2 family. {ECO:0000305}.
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DR EMBL; AF284574; AAF87103.1; -; mRNA.
DR EMBL; AF260334; AAG44665.1; -; mRNA.
DR EMBL; AK313704; BAG36450.1; -; mRNA.
DR EMBL; CH471052; EAW79824.1; -; Genomic_DNA.
DR EMBL; CH471052; EAW79825.1; -; Genomic_DNA.
DR EMBL; BC020620; AAH20620.1; -; mRNA.
DR EMBL; BC107890; AAI07891.1; -; mRNA.
DR CCDS; CCDS33806.1; -.
DR RefSeq; NP_064587.1; NM_020202.4.
DR AlphaFoldDB; Q9NQR4; -.
DR SMR; Q9NQR4; -.
DR BioGRID; 121278; 48.
DR IntAct; Q9NQR4; 7.
DR MINT; Q9NQR4; -.
DR STRING; 9606.ENSP00000377696; -.
DR iPTMnet; Q9NQR4; -.
DR PhosphoSitePlus; Q9NQR4; -.
DR BioMuta; NIT2; -.
DR DMDM; 74725271; -.
DR REPRODUCTION-2DPAGE; IPI00549467; -.
DR UCD-2DPAGE; Q9NQR4; -.
DR CPTAC; CPTAC-549; -.
DR CPTAC; CPTAC-550; -.
DR EPD; Q9NQR4; -.
DR jPOST; Q9NQR4; -.
DR MassIVE; Q9NQR4; -.
DR PaxDb; Q9NQR4; -.
DR PeptideAtlas; Q9NQR4; -.
DR PRIDE; Q9NQR4; -.
DR ProteomicsDB; 82173; -.
DR Antibodypedia; 32221; 256 antibodies from 24 providers.
DR DNASU; 56954; -.
DR Ensembl; ENST00000394140.9; ENSP00000377696.3; ENSG00000114021.12.
DR GeneID; 56954; -.
DR KEGG; hsa:56954; -.
DR MANE-Select; ENST00000394140.9; ENSP00000377696.3; NM_020202.5; NP_064587.1.
DR CTD; 56954; -.
DR DisGeNET; 56954; -.
DR GeneCards; NIT2; -.
DR HGNC; HGNC:29878; NIT2.
DR HPA; ENSG00000114021; Tissue enhanced (liver).
DR neXtProt; NX_Q9NQR4; -.
DR OpenTargets; ENSG00000114021; -.
DR PharmGKB; PA134882857; -.
DR VEuPathDB; HostDB:ENSG00000114021; -.
DR eggNOG; KOG0806; Eukaryota.
DR GeneTree; ENSGT00550000074838; -.
DR HOGENOM; CLU_030130_1_0_1; -.
DR InParanoid; Q9NQR4; -.
DR OMA; GNTYRES; -.
DR OrthoDB; 1154369at2759; -.
DR PhylomeDB; Q9NQR4; -.
DR TreeFam; TF300747; -.
DR BRENDA; 3.5.1.3; 2681.
DR PathwayCommons; Q9NQR4; -.
DR Reactome; R-HSA-6798695; Neutrophil degranulation.
DR SignaLink; Q9NQR4; -.
DR BioGRID-ORCS; 56954; 57 hits in 1081 CRISPR screens.
DR ChiTaRS; NIT2; human.
DR GenomeRNAi; 56954; -.
DR Pharos; Q9NQR4; Tbio.
DR PRO; PR:Q9NQR4; -.
DR Proteomes; UP000005640; Chromosome 3.
DR RNAct; Q9NQR4; protein.
DR Bgee; ENSG00000114021; Expressed in right lobe of liver and 200 other tissues.
DR ExpressionAtlas; Q9NQR4; baseline and differential.
DR Genevisible; Q9NQR4; HS.
DR GO; GO:0005813; C:centrosome; IDA:HPA.
DR GO; GO:0005829; C:cytosol; IDA:HPA.
DR GO; GO:0070062; C:extracellular exosome; HDA:UniProtKB.
DR GO; GO:0005576; C:extracellular region; TAS:Reactome.
DR GO; GO:0035580; C:specific granule lumen; TAS:Reactome.
DR GO; GO:1904724; C:tertiary granule lumen; TAS:Reactome.
DR GO; GO:0106008; F:2-oxoglutaramate amidase activity; IEA:RHEA.
DR GO; GO:0050152; F:omega-amidase activity; IDA:BHF-UCL.
DR GO; GO:0006528; P:asparagine metabolic process; IDA:BHF-UCL.
DR GO; GO:0006541; P:glutamine metabolic process; IDA:BHF-UCL.
DR GO; GO:0006107; P:oxaloacetate metabolic process; IDA:BHF-UCL.
DR CDD; cd07572; nit; 1.
DR Gene3D; 3.60.110.10; -; 1.
DR InterPro; IPR003010; C-N_Hydrolase.
DR InterPro; IPR036526; C-N_Hydrolase_sf.
DR InterPro; IPR045254; Nit1/2_C-N_Hydrolase.
DR Pfam; PF00795; CN_hydrolase; 1.
DR SUPFAM; SSF56317; SSF56317; 1.
DR PROSITE; PS50263; CN_HYDROLASE; 1.
PE 1: Evidence at protein level;
KW Acetylation; Cytoplasm; Hydrolase; Phosphoprotein; Reference proteome.
FT CHAIN 1..276
FT /note="Omega-amidase NIT2"
FT /id="PRO_0000320252"
FT DOMAIN 4..248
FT /note="CN hydrolase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT ACT_SITE 43
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054,
FT ECO:0000305|PubMed:22674578"
FT ACT_SITE 112
FT /note="Proton donor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054,
FT ECO:0000305|PubMed:22674578"
FT ACT_SITE 153
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054,
FT ECO:0000305|PubMed:22674578"
FT MOD_RES 26
FT /note="Phosphoserine"
FT /evidence="ECO:0007744|PubMed:24275569"
FT MOD_RES 68
FT /note="N6-acetyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q9JHW2"
FT MOD_RES 68
FT /note="N6-succinyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q9JHW2"
FT MOD_RES 123
FT /note="N6-succinyllysine"
FT /evidence="ECO:0000250|UniProtKB:Q9JHW2"
FT MOD_RES 130
FT /note="N6-succinyllysine"
FT /evidence="ECO:0000250|UniProtKB:Q9JHW2"
FT VARIANT 231
FT /note="V -> A (in dbSNP:rs17851799)"
FT /evidence="ECO:0000269|PubMed:15489334"
FT /id="VAR_039180"
FT MUTAGEN 43
FT /note="E->A: Loss of activity using succinamate as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22674578"
FT MUTAGEN 112
FT /note="K->A: Loss of activity using succinamate as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22674578"
FT MUTAGEN 116..128
FT /note="Missing: Less than 3% of wild-type activity using
FT succinamate as substrate."
FT /evidence="ECO:0000269|PubMed:22674578"
FT MUTAGEN 153
FT /note="C->A: Loss of activity using succinamate as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22674578"
SQ SEQUENCE 276 AA; 30608 MW; 32FA797601A419C6 CRC64;
MTSFRLALIQ LQISSIKSDN VTRACSFIRE AATQGAKIVS LPECFNSPYG AKYFPEYAEK
IPGESTQKLS EVAKECSIYL IGGSIPEEDA GKLYNTCAVF GPDGTLLAKY RKIHLFDIDV
PGKITFQESK TLSPGDSFST FDTPYCRVGL GICYDMRFAE LAQIYAQRGC QLLVYPGAFN
LTTGPAHWEL LQRSRAVDNQ VYVATASPAR DDKASYVAWG HSTVVNPWGE VLAKAGTEEA
IVYSDIDLKK LAEIRQQIPV FRQKRSDLYA VEMKKP
