PNCA_MYCTU
ID PNCA_MYCTU Reviewed; 186 AA.
AC I6XD65;
DT 18-JAN-2017, integrated into UniProtKB/Swiss-Prot.
DT 03-OCT-2012, sequence version 1.
DT 03-AUG-2022, entry version 67.
DE RecName: Full=Nicotinamidase/pyrazinamidase {ECO:0000303|PubMed:18201201};
DE AltName: Full=Nicotinamidase {ECO:0000303|PubMed:18201201};
DE EC=3.5.1.19 {ECO:0000269|PubMed:18201201};
DE AltName: Full=Nicotinamide deamidase {ECO:0000250|UniProtKB:P21369};
DE Short=NAMase {ECO:0000250|UniProtKB:P21369};
DE AltName: Full=Pyrazinamidase {ECO:0000303|PubMed:18201201, ECO:0000303|PubMed:21283666};
DE Short=PZAase {ECO:0000250|UniProtKB:P21369};
DE EC=3.5.1.- {ECO:0000269|PubMed:18201201};
GN Name=pncA {ECO:0000303|PubMed:18201201, ECO:0000312|EMBL:CCP44816.1};
GN OrderedLocusNames=Rv2043c {ECO:0000312|EMBL:CCP44816.1},
GN LH57_11125 {ECO:0000312|EMBL:AIR14792.1};
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 NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27294 / TMC 102 / H37Rv;
RA Hazbon M.H., Riojas M.A., Damon A.M., Alalade R.O., Cantwell B.J.,
RA Monaco A., King S., Sohrabi A.;
RT "Phylogenetic analysis of Mycobacterial species using whole genome
RT sequences.";
RL Submitted (SEP-2014) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP ACTIVITY REGULATION, SUBUNIT, AND MUTAGENESIS OF ASP-8; ASP-49; HIS-51;
RP HIS-57; SER-59; HIS-71; LYS-96; SER-104 AND CYS-138.
RC STRAIN=H37Rv;
RX PubMed=18201201; DOI=10.1111/j.1742-4658.2007.06241.x;
RA Zhang H., Deng J.Y., Bi L.J., Zhou Y.F., Zhang Z.P., Zhang C.G., Zhang Y.,
RA Zhang X.E.;
RT "Characterization of Mycobacterium tuberculosis
RT nicotinamidase/pyrazinamidase.";
RL FEBS J. 275:753-762(2008).
RN [4]
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).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH IRON, COFACTOR,
RP REACTION MECHANISM, AND ACTIVE SITE.
RC STRAIN=H37Rv;
RX PubMed=21283666; DOI=10.1371/journal.pone.0015785;
RA Petrella S., Gelus-Ziental N., Maudry A., Laurans C., Boudjelloul R.,
RA Sougakoff W.;
RT "Crystal structure of the pyrazinamidase of Mycobacterium tuberculosis:
RT insights into natural and acquired resistance to pyrazinamide.";
RL PLoS ONE 6:E15785-E15785(2011).
CC -!- FUNCTION: Catalyzes the deamidation of nicotinamide (NAM) into
CC nicotinate (PubMed:18201201). Likely functions in the cyclical salvage
CC pathway for production of NAD from nicotinamide (By similarity).
CC {ECO:0000250|UniProtKB:P21369, ECO:0000269|PubMed:18201201}.
CC -!- FUNCTION: Is involved in the activation of the first-line
CC antituberculous drug pyrazinamide (PZA) by converting it into the
CC active form, pyrazinoic acid. {ECO:0000269|PubMed:18201201}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + nicotinamide = NH4(+) + nicotinate;
CC Xref=Rhea:RHEA:14545, ChEBI:CHEBI:15377, ChEBI:CHEBI:17154,
CC ChEBI:CHEBI:28938, ChEBI:CHEBI:32544; EC=3.5.1.19;
CC Evidence={ECO:0000269|PubMed:18201201};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + pyrazinamide = NH4(+) + pyrazine-2-carboxylate;
CC Xref=Rhea:RHEA:35063, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:45285, ChEBI:CHEBI:71266;
CC Evidence={ECO:0000269|PubMed:18201201};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:18201201};
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:18201201, ECO:0000269|PubMed:21283666};
CC Note=PncA contains Mn(2+) and Fe(2+) in a molecular ratio of 1:1. PncA
CC has only one metal center. It is believed that PncA binds iron in the
CC natural state, although both metals can support enzymatic activity.
CC {ECO:0000269|PubMed:18201201};
CC -!- ACTIVITY REGULATION: Is inhibited by Cu(2+), Zn(2+) and Fe(3+).
CC {ECO:0000269|PubMed:18201201}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7.0 for the hydrolysis of nicotinamide. The enzyme
CC activity decreases rapidly below pH 6.0 or above pH 8.0.
CC {ECO:0000269|PubMed:18201201};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius for the hydrolysis of
CC nicotinamide. Below 25 or above 70 degrees Celsius, the enzyme loses
CC its activity rapidly. {ECO:0000269|PubMed:18201201};
CC -!- PATHWAY: Cofactor biosynthesis; nicotinate biosynthesis; nicotinate
CC from nicotinamide: step 1/1. {ECO:0000250|UniProtKB:P21369}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:18201201}.
CC -!- MISCELLANEOUS: Most pyrazinamide-resistant clinical M.tuberculosis
CC isolates contain mutations in the pncA gene, causing lack or reduction
CC of PZAase activity. {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the isochorismatase family. {ECO:0000305}.
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DR EMBL; AL123456; CCP44816.1; -; Genomic_DNA.
DR EMBL; CP009480; AIR14792.1; -; Genomic_DNA.
DR RefSeq; NP_216559.1; NC_000962.3.
DR RefSeq; WP_003410243.1; NZ_NVQJ01000046.1.
DR PDB; 3PL1; X-ray; 2.20 A; A=1-186.
DR PDBsum; 3PL1; -.
DR AlphaFoldDB; I6XD65; -.
DR SMR; I6XD65; -.
DR STRING; 83332.Rv2043c; -.
DR PaxDb; I6XD65; -.
DR DNASU; 888260; -.
DR GeneID; 45426023; -.
DR GeneID; 888260; -.
DR KEGG; mtu:Rv2043c; -.
DR PATRIC; fig|83332.111.peg.2278; -.
DR TubercuList; Rv2043c; -.
DR eggNOG; COG1335; Bacteria.
DR HOGENOM; CLU_068979_13_2_11; -.
DR OMA; DFVDSWP; -.
DR PhylomeDB; I6XD65; -.
DR BRENDA; 3.5.1.19; 3445.
DR BRENDA; 3.5.1.B15; 3445.
DR UniPathway; UPA00830; UER00790.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0008198; F:ferrous iron binding; IDA:UniProtKB.
DR GO; GO:0016811; F:hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in linear amides; IBA:GO_Central.
DR GO; GO:0030145; F:manganese ion binding; IDA:UniProtKB.
DR GO; GO:0008936; F:nicotinamidase activity; IDA:UniProtKB.
DR GO; GO:0006769; P:nicotinamide metabolic process; IDA:UniProtKB.
DR GO; GO:0019363; P:pyridine nucleotide biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR GO; GO:0006805; P:xenobiotic metabolic process; IDA:UniProtKB.
DR Gene3D; 3.40.50.850; -; 1.
DR InterPro; IPR000868; Isochorismatase-like.
DR InterPro; IPR036380; Isochorismatase-like_sf.
DR Pfam; PF00857; Isochorismatase; 1.
DR SUPFAM; SSF52499; SSF52499; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Hydrolase; Iron; Manganese;
KW Metal-binding; Pyridine nucleotide biosynthesis; Reference proteome.
FT CHAIN 1..186
FT /note="Nicotinamidase/pyrazinamidase"
FT /id="PRO_0000438715"
FT ACT_SITE 8
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:21283666"
FT ACT_SITE 96
FT /evidence="ECO:0000305|PubMed:21283666"
FT ACT_SITE 138
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:21283666"
FT BINDING 49
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:21283666,
FT ECO:0007744|PDB:3PL1"
FT BINDING 51
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:21283666,
FT ECO:0007744|PDB:3PL1"
FT BINDING 57
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:21283666,
FT ECO:0007744|PDB:3PL1"
FT BINDING 71
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:21283666,
FT ECO:0007744|PDB:3PL1"
FT MUTAGEN 8
FT /note="D->A: Loss of enzymatic activity. No effect on metal
FT ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 49
FT /note="D->A: 3000-fold decrease in enzymatic activity with
FT NAM as substrate, and 410-fold decrease in enzymatic
FT activity with PZA as substrate. The mutant has no
FT detectable iron and manganese."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 51
FT /note="H->A: 10-fold decrease in enzymatic activity with
FT NAM as substrate, and 21-fold decrease in enzymatic
FT activity with PZA as substrate. The mutant has a low metal
FT ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 57
FT /note="H->A: 112-fold decrease in enzymatic activity with
FT NAM as substrate, and 164-fold decrease in enzymatic
FT activity with PZA as substrate. The mutant has no
FT detectable iron and manganese."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 59
FT /note="S->A: 2.4-fold decrease in enzymatic activity with
FT NAM or PZA as substrate. No effect on metal ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 71
FT /note="H->A: 100-fold decrease in enzymatic activity with
FT NAM or PZA as substrate. The mutant has a low metal ion
FT content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 96
FT /note="K->A: Loss of enzymatic activity. No effect on metal
FT ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 104
FT /note="S->A: 5-fold decrease in enzymatic activity with NAM
FT as substrate, and 3-fold decrease in enzymatic activity
FT with PZA as substrate. No effect on metal ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
FT MUTAGEN 138
FT /note="C->A: Loss of enzymatic activity. No effect on metal
FT ion content."
FT /evidence="ECO:0000269|PubMed:18201201"
SQ SEQUENCE 186 AA; 19605 MW; B303D652A263A4F5 CRC64;
MRALIIVDVQ NDFCEGGSLA VTGGAALARA ISDYLAEAAD YHHVVATKDF HIDPGDHFSG
TPDYSSSWPP HCVSGTPGAD FHPSLDTSAI EAVFYKGAYT GAYSGFEGVD ENGTPLLNWL
RQRGVDEVDV VGIATDHCVR QTAEDAVRNG LATRVLVDLT AGVSADTTVA ALEEMRTASV
ELVCSS
