ASPG_MYCTU
ID ASPG_MYCTU Reviewed; 315 AA.
AC P9WPX5; L0T751; P63627; Q10759;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 40.
DE RecName: Full=L-asparaginase {ECO:0000303|PubMed:33412160};
DE Short=L-ASNase;
DE EC=3.5.1.1 {ECO:0000269|PubMed:24586151, ECO:0000269|PubMed:33412160};
DE AltName: Full=L-asparagine amidohydrolase;
GN Name=ansA {ECO:0000303|PubMed:24586151}; OrderedLocusNames=Rv1538c;
GN ORFNames=MTCY48.27;
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 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 [3]
RP FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, AND DISRUPTION
RP PHENOTYPE.
RX PubMed=24586151; DOI=10.1371/journal.ppat.1003928;
RA Gouzy A., Larrouy-Maumus G., Bottai D., Levillain F., Dumas A.,
RA Wallach J.B., Caire-Brandli I., de Chastellier C., Wu T.D., Poincloux R.,
RA Brosch R., Guerquin-Kern J.L., Schnappinger D., Sorio de Carvalho L.P.,
RA Poquet Y., Neyrolles O.;
RT "Mycobacterium tuberculosis exploits asparagine to assimilate nitrogen and
RT resist acid stress during infection.";
RL PLoS Pathog. 10:e1003928-e1003928(2014).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, AND IDENTIFICATION BY MASS SPECTROMETRY.
RC STRAIN=CCDC 5079;
RX PubMed=33412160; DOI=10.1016/j.biochi.2020.12.023;
RA Kataria A., Patel A.K., Kundu B.;
RT "Distinct functional properties of secretory L-asparaginase Rv1538c
RT involved in phagosomal survival of Mycobacterium tuberculosis.";
RL Biochimie 182:1-12(2021).
CC -!- FUNCTION: Has a dual function in both nitrogen assimilation and in
CC protection against acid stress during infection through asparagine
CC hydrolysis and NH4(+) release (PubMed:24586151). Catalyzes asparagine
CC hydrolysis (PubMed:24586151, PubMed:33412160). Cannot use glutamine
CC (PubMed:24586151, PubMed:33412160). Required for intracellular survival
CC inside macrophages during host colonization (PubMed:24586151). Mediates
CC phagosome acidification arrest and resistance to acid stress through
CC the formation of acid-neutralizing NH4(+) ions (PubMed:24586151). In
CC addition, may induce stress to primary immune cells and compromise the
CC host immune response (PubMed:33412160). {ECO:0000269|PubMed:24586151,
CC ECO:0000269|PubMed:33412160}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-asparagine = L-aspartate + NH4(+);
CC Xref=Rhea:RHEA:21016, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:29991, ChEBI:CHEBI:58048; EC=3.5.1.1;
CC Evidence={ECO:0000269|PubMed:24586151, ECO:0000269|PubMed:33412160};
CC -!- ACTIVITY REGULATION: Activity in enhanced in the presence of K(+) and
CC Mn(2+) and reduced by about 40% in the presence of Cu(2+).
CC {ECO:0000269|PubMed:33412160}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8.3 mM for L-asparagine (at pH 8.0) {ECO:0000269|PubMed:33412160};
CC KM=19.57 mM for L-asparagine (at pH 7.4)
CC {ECO:0000269|PubMed:33412160};
CC KM=0.38 mM for L-asparagine (at pH 5.5)
CC {ECO:0000269|PubMed:33412160};
CC Vmax=797 umol/min/mg enzyme (at pH 8.0)
CC {ECO:0000269|PubMed:33412160};
CC Vmax=1587 umol/min/mg enzyme (at pH 7.4)
CC {ECO:0000269|PubMed:33412160};
CC Vmax=74 umol/min/mg enzyme (at pH 5.5) {ECO:0000269|PubMed:33412160};
CC Note=kcat is 869 sec(-1) (at pH 8.0). kcat is 1732 sec(-1) (at pH
CC 7.4). kcat is 80.7 sec(-1) (at pH 5.5).
CC {ECO:0000269|PubMed:33412160};
CC pH dependence:
CC Optimum pH is 8.0. Retains about 40% activity at pH 5.5.
CC {ECO:0000269|PubMed:33412160};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius. Shows maximal activity
CC between 30 and 40 degrees Celsius. {ECO:0000269|PubMed:33412160};
CC -!- SUBUNIT: Homodimer (PubMed:33412160). Can form tetramers and higher
CC multimers, probably resulting from the dynamic association of dimers
CC (PubMed:33412160). {ECO:0000269|PubMed:33412160}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:24586151}.
CC Note=Probably secreted via the ESX-5 / type VII secretion system
CC (T7SS). Secreted in host cells during infection.
CC {ECO:0000269|PubMed:24586151}.
CC -!- DISRUPTION PHENOTYPE: Not essential. The growth of the knockout mutant
CC is impaired, although not fully abolished, when asparagine is provided
CC as sole nitrogen donor. Nitrogen assimilation from asparagine into
CC glutamate and glutamine is fully abrogated in the knockout mutant at
CC acidic pH. Mutant is impaired in host tissue colonization.
CC {ECO:0000269|PubMed:24586151}.
CC -!- SIMILARITY: Belongs to the asparaginase 1 family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CCP44302.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; AL123456; CCP44302.1; ALT_INIT; Genomic_DNA.
DR PIR; G70760; G70760.
DR RefSeq; NP_216054.1; NC_000962.3.
DR AlphaFoldDB; P9WPX5; -.
DR SMR; P9WPX5; -.
DR STRING; 83332.Rv1538c; -.
DR PaxDb; P9WPX5; -.
DR DNASU; 886410; -.
DR GeneID; 886410; -.
DR KEGG; mtu:Rv1538c; -.
DR TubercuList; Rv1538c; -.
DR eggNOG; COG0252; Bacteria.
DR OMA; RKNHTSR; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0042597; C:periplasmic space; IBA:GO_Central.
DR GO; GO:0004067; F:asparaginase activity; IBA:GO_Central.
DR GO; GO:0006530; P:asparagine catabolic process; IBA:GO_Central.
DR CDD; cd08964; L-asparaginase_II; 1.
DR Gene3D; 3.40.50.1170; -; 1.
DR Gene3D; 3.40.50.40; -; 1.
DR InterPro; IPR004550; AsnASE_II.
DR InterPro; IPR036152; Asp/glu_Ase-like_sf.
DR InterPro; IPR006034; Asparaginase/glutaminase-like.
DR InterPro; IPR020827; Asparaginase/glutaminase_AS1.
DR InterPro; IPR027475; Asparaginase/glutaminase_AS2.
DR InterPro; IPR040919; Asparaginase_C.
DR InterPro; IPR027473; L-asparaginase_C.
DR InterPro; IPR027474; L-asparaginase_N.
DR InterPro; IPR037152; L-asparaginase_N_sf.
DR Pfam; PF00710; Asparaginase; 1.
DR Pfam; PF17763; Asparaginase_C; 1.
DR PIRSF; PIRSF001220; L-ASNase_gatD; 1.
DR PRINTS; PR00139; ASNGLNASE.
DR SMART; SM00870; Asparaginase; 1.
DR SUPFAM; SSF53774; SSF53774; 1.
DR PROSITE; PS00144; ASN_GLN_ASE_1; 1.
DR PROSITE; PS00917; ASN_GLN_ASE_2; 1.
DR PROSITE; PS51732; ASN_GLN_ASE_3; 1.
PE 1: Evidence at protein level;
KW Hydrolase; Reference proteome; Secreted.
FT CHAIN 1..315
FT /note="L-asparaginase"
FT /id="PRO_0000171084"
FT DOMAIN 2..315
FT /note="Asparaginase/glutaminase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01068"
FT ACT_SITE 12
FT /note="O-isoaspartyl threonine intermediate"
FT /evidence="ECO:0000250|UniProtKB:P00805"
FT BINDING 52..53
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P00805"
FT BINDING 84..85
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P00805"
SQ SEQUENCE 315 AA; 31741 MW; AEC46476F73FA340 CRC64;
MARLTVITTG GTISTTAGPD GVLRPTHCGA TLIAGLDMDS DIEVVDLMAL DSSKLTPADW
DRIGAAVQEA FRGGADGVVI THGTDTLEET ALWLDLTYAG SRPVVLTGAM LSADAPGADG
PANLRDALAV AADPAARDLG VLVSFGGRVL QPLGLHKVAN PDLCGFAGES LGFTSGGVRL
TRTKTRPYLG DLGAAVAPRV DIVAVYPGSD AVAMDACVAA GARAVVLEAL GSGNAGAAVI
EGVRRHCRDG SDPVVIAVST RVAGARVGAG YGPGHDLVEA GAVMVPRLPP SQARVLLMAA
LAANSPVADV IDRWG
