DHA_MYCTU
ID DHA_MYCTU Reviewed; 371 AA.
AC P9WQB1; L0TC82; O33322; P30234;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 43.
DE RecName: Full=Alanine dehydrogenase;
DE EC=1.4.1.1 {ECO:0000269|PubMed:10527947, ECO:0000269|PubMed:18304579};
DE AltName: Full=40 kDa antigen;
DE AltName: Full=TB43;
GN Name=ald; OrderedLocusNames=Rv2780; ORFNames=MTV002.45;
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 [GENOMIC DNA].
RC STRAIN=ATCC 35801 / TMC 107 / Erdman;
RX PubMed=1587598; DOI=10.1128/iai.60.6.2317-2323.1992;
RA Andersen A.B., Andersen P., Ljungqvist L.;
RT "Structure and function of a 40,000-molecular-weight protein antigen of
RT Mycobacterium tuberculosis.";
RL Infect. Immun. 60:2317-2323(1992).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RA Singh M., Hutter B.;
RT "Host-vector system for high level expression and purification of
RT enzymatically active L-alanine dehydrogenase of M.tuberculosis in E.coli.";
RL Submitted (MAR-1997) to the EMBL/GenBank/DDBJ databases.
RN [3]
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 [4]
RP PROTEIN SEQUENCE OF 1-21.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=7822223; DOI=10.1111/j.1365-2672.1994.tb02813.x;
RA Deshpande R.G., Khan M.B., Bhat D.A., Navalkar R.G.;
RT "Isolation of a 43 kDa protein from Mycobacterium tuberculosis H37Rv and
RT its identification as a pyridine nucleotide transhydrogenase.";
RL J. Appl. Bacteriol. 77:639-643(1994).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP SUBCELLULAR LOCATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=10527947; DOI=10.1042/bj3430669;
RA Hutter B., Singh M.;
RT "Properties of the 40 kDa antigen of Mycobacterium tuberculosis, a
RT functional L-alanine dehydrogenase.";
RL Biochem. J. 343:669-672(1999).
RN [6]
RP INDUCTION BY STARVATION.
RX PubMed=11929527; DOI=10.1046/j.1365-2958.2002.02779.x;
RA Betts J.C., Lukey P.T., Robb L.C., McAdam R.A., Duncan K.;
RT "Evaluation of a nutrient starvation model of Mycobacterium tuberculosis
RT persistence by gene and protein expression profiling.";
RL Mol. Microbiol. 43:717-731(2002).
RN [7]
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 [8]
RP X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF WILD TYPE AND OF MUTANT ASN-270 IN
RP COMPLEX WITH NAD AND SUBSTRATE ANALOGS, MUTAGENESIS OF HIS-96 AND ASP-270,
RP ACTIVE SITE, BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM, CATALYTIC
RP ACTIVITY, COFACTOR, AND SUBUNIT.
RX PubMed=18304579; DOI=10.1016/j.jmb.2008.01.091;
RA Agren D., Stehr M., Berthold C.L., Kapoor S., Oehlmann W., Singh M.,
RA Schneider G.;
RT "Three-dimensional structures of apo- and holo-L-alanine dehydrogenase from
RT Mycobacterium tuberculosis reveal conformational changes upon coenzyme
RT binding.";
RL J. Mol. Biol. 377:1161-1173(2008).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) IN COMPLEX WITH NAD AND SUBSTRATE
RP ANALOGS, REACTION MECHANISM, AND SUBUNIT.
RX PubMed=18491387; DOI=10.1002/prot.22101;
RA Tripathi S.M., Ramachandran R.;
RT "Crystal structures of the Mycobacterium tuberculosis secretory antigen
RT alanine dehydrogenase (Rv2780) in apo and ternary complex forms captures
RT 'open' and 'closed' enzyme conformations.";
RL Proteins 72:1089-1095(2008).
CC -!- FUNCTION: Catalyzes the reversible reductive amination of pyruvate to
CC L-alanine. However, since the physiological environment of
CC M.tuberculosis has a neutral pH, it can be assumed that the enzyme
CC catalyzes exclusively the formation of L-alanine. May play a role in
CC cell wall synthesis as L-alanine is an important constituent of the
CC peptidoglycan layer. {ECO:0000269|PubMed:10527947}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-alanine + NAD(+) = H(+) + NADH + NH4(+) + pyruvate;
CC Xref=Rhea:RHEA:18405, ChEBI:CHEBI:15361, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:28938, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:57972; EC=1.4.1.1;
CC Evidence={ECO:0000269|PubMed:10527947, ECO:0000269|PubMed:18304579};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:18304579};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000269|PubMed:18304579};
CC -!- ACTIVITY REGULATION: Inhibited by CuSO(4) and ZnCl(2).
CC {ECO:0000269|PubMed:10527947}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=98.2 uM for NADH (at pH 7.4 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10527947};
CC KM=0.31 mM for NAD (at pH 10.2 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10527947};
CC KM=0.76 mM for pyruvate (at pH 7.4 and at 25 degrees Celsius)
CC {ECO:0000269|PubMed:18304579};
CC KM=1.45 mM for pyruvate (at pH 7.4 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10527947};
CC KM=13.8 mM for L-alanine (at pH 10.2 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10527947};
CC KM=15.6 mM for L-alanine (at pH 10.2 and at 25 degrees Celsius)
CC {ECO:0000269|PubMed:18304579};
CC KM=35.4 mM for ammonium (at pH 7.4 and at 37 degrees Celsius)
CC {ECO:0000269|PubMed:10527947};
CC Vmax=31.8 umol/min/mg enzyme for reductive amination
CC {ECO:0000269|PubMed:10527947};
CC Vmax=23.7 umol/min/mg enzyme for oxidative deamination
CC {ECO:0000269|PubMed:10527947};
CC pH dependence:
CC Optimum pH is between 10 and 11 for the oxidative deamination and
CC between 7 and 7.5 for the reductive amination.
CC {ECO:0000269|PubMed:10527947};
CC Temperature dependence:
CC Relatively stable, it loses only 25% of its total activity after 4
CC hours at 60 degrees Celsius. The enzyme is much more active at
CC temperatures above 37 degrees Celsius, particularly in the reductive
CC amination. The velocity almost doubles at temperatures between 60-65
CC degrees Celsius compared with 37 degrees Celsius. Above 65 degrees
CC Celsius there is a sharp decrease in activity, due to thermal
CC inactivation of the enzyme. {ECO:0000269|PubMed:10527947,
CC ECO:0000269|PubMed:18304579};
CC -!- PATHWAY: Amino-acid degradation; L-alanine degradation via
CC dehydrogenase pathway; NH(3) and pyruvate from L-alanine: step 1/1.
CC -!- SUBUNIT: Homohexamer. Trimer of dimers. {ECO:0000269|PubMed:18304579,
CC ECO:0000269|PubMed:18491387}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:10527947}.
CC -!- INDUCTION: Upon nutrient starvation. {ECO:0000269|PubMed:11929527}.
CC -!- MISCELLANEOUS: L-alanine dehydrogenase activity increases when the
CC M.tuberculosis is shifted from aerobic to anaerobic growth conditions.
CC {ECO:0000305|PubMed:18491387}.
CC -!- SIMILARITY: Belongs to the AlaDH/PNT family. {ECO:0000305}.
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DR EMBL; X63069; CAA44791.1; -; Genomic_DNA.
DR EMBL; U92472; AAC38804.1; -; Genomic_DNA.
DR EMBL; AL123456; CCP45579.1; -; Genomic_DNA.
DR PIR; C70883; A43830.
DR RefSeq; NP_217296.1; NC_000962.3.
DR RefSeq; WP_003899477.1; NZ_NVQJ01000020.1.
DR PDB; 2VHV; X-ray; 2.80 A; A/B=1-371.
DR PDB; 2VHW; X-ray; 2.00 A; A/B/C/D/E/F=1-371.
DR PDB; 2VHX; X-ray; 2.00 A; A/B/C/D/E/F=1-371.
DR PDB; 2VHY; X-ray; 2.30 A; A/B=1-371.
DR PDB; 2VHZ; X-ray; 2.04 A; A/B=1-371.
DR PDB; 2VOE; X-ray; 2.60 A; A/B/C/D/E/F=1-371.
DR PDB; 2VOJ; X-ray; 2.60 A; A/C/E=1-371.
DR PDB; 6O7F; X-ray; 2.30 A; A=1-371.
DR PDBsum; 2VHV; -.
DR PDBsum; 2VHW; -.
DR PDBsum; 2VHX; -.
DR PDBsum; 2VHY; -.
DR PDBsum; 2VHZ; -.
DR PDBsum; 2VOE; -.
DR PDBsum; 2VOJ; -.
DR PDBsum; 6O7F; -.
DR AlphaFoldDB; P9WQB1; -.
DR SMR; P9WQB1; -.
DR STRING; 83332.Rv2780; -.
DR PaxDb; P9WQB1; -.
DR DNASU; 888493; -.
DR GeneID; 45426769; -.
DR GeneID; 888493; -.
DR KEGG; mtu:Rv2780; -.
DR TubercuList; Rv2780; -.
DR eggNOG; COG0686; Bacteria.
DR OMA; HPYYHLY; -.
DR PhylomeDB; P9WQB1; -.
DR BioCyc; MetaCyc:G185E-7029-MON; -.
DR BRENDA; 1.4.1.1; 3445.
DR UniPathway; UPA00527; UER00585.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005829; C:cytosol; HDA:MTBBASE.
DR GO; GO:0005576; C:extracellular region; IDA:MTBBASE.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; HDA:MTBBASE.
DR GO; GO:0005886; C:plasma membrane; HDA:MTBBASE.
DR GO; GO:0000286; F:alanine dehydrogenase activity; IDA:MTBBASE.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0006524; P:alanine catabolic process; IDA:MTBBASE.
DR GO; GO:0042853; P:L-alanine catabolic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0001666; P:response to hypoxia; IEP:MTBBASE.
DR CDD; cd05305; L-AlaDH; 1.
DR InterPro; IPR008141; Ala_DH.
DR InterPro; IPR008143; Ala_DH/PNT_CS2.
DR InterPro; IPR008142; AlaDH/PNT_CS1.
DR InterPro; IPR007886; AlaDH/PNT_N.
DR InterPro; IPR007698; AlaDH/PNT_NAD(H)-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR PANTHER; PTHR42795; PTHR42795; 1.
DR Pfam; PF01262; AlaDh_PNT_C; 1.
DR Pfam; PF05222; AlaDh_PNT_N; 1.
DR PIRSF; PIRSF000183; Alanine_dh; 1.
DR SMART; SM01002; AlaDh_PNT_C; 1.
DR SMART; SM01003; AlaDh_PNT_N; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR00518; alaDH; 1.
DR PROSITE; PS00836; ALADH_PNT_1; 1.
DR PROSITE; PS00837; ALADH_PNT_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Magnesium; Metal-binding; NAD;
KW Nucleotide-binding; Oxidoreductase; Reference proteome; Secreted.
FT CHAIN 1..371
FT /note="Alanine dehydrogenase"
FT /id="PRO_0000198994"
FT ACT_SITE 96
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000269|PubMed:18304579"
FT ACT_SITE 270
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000269|PubMed:18304579"
FT BINDING 15
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:18304579"
FT BINDING 75
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:18304579"
FT BINDING 134
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 178..179
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 198
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 203
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 220
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 239..240
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 267..270
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 279
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 298..301
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0000269|PubMed:18491387"
FT BINDING 323
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0007744|PDB:2VHX"
FT BINDING 327
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:18304579,
FT ECO:0007744|PDB:2VHX"
FT MUTAGEN 96
FT /note="H->A: Completely inactive mutant."
FT /evidence="ECO:0000269|PubMed:18304579"
FT MUTAGEN 270
FT /note="D->A: Completely inactive mutant."
FT /evidence="ECO:0000269|PubMed:18304579"
FT MUTAGEN 270
FT /note="D->N: Completely inactive mutant. The bifurcated
FT hydrogen bond between D-270 and the ribose of NAD is
FT replaced by a single hydrogen bond."
FT /evidence="ECO:0000269|PubMed:18304579"
FT CONFLICT 13
FT /note="E -> EFQ (in Ref. 1; CAA44791)"
FT /evidence="ECO:0000305"
FT STRAND 2..5
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 20..28
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 32..36
FT /evidence="ECO:0007829|PDB:2VHW"
FT TURN 37..40
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 41..43
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 47..53
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 56..59
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 61..67
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 69..72
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 79..84
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 90..93
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 97..99
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 101..110
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 113..116
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 117..119
FT /evidence="ECO:0007829|PDB:2VHW"
FT TURN 128..130
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 131..148
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 151..153
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 170..174
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 178..189
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 193..199
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 201..210
FT /evidence="ECO:0007829|PDB:2VHW"
FT TURN 211..213
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 214..219
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 222..231
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 233..237
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 242..244
FT /evidence="ECO:0007829|PDB:6O7F"
FT HELIX 252..255
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 263..266
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 267..270
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 283..285
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 287..290
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 293..296
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 298..300
FT /evidence="ECO:0007829|PDB:6O7F"
FT HELIX 301..304
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 306..335
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 337..340
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 343..346
FT /evidence="ECO:0007829|PDB:2VHW"
FT HELIX 353..359
FT /evidence="ECO:0007829|PDB:2VHW"
FT STRAND 360..362
FT /evidence="ECO:0007829|PDB:2VOJ"
FT HELIX 367..370
FT /evidence="ECO:0007829|PDB:2VHW"
SQ SEQUENCE 371 AA; 38713 MW; 9DF7540524DC116A CRC64;
MRVGIPTETK NNEFRVAITP AGVAELTRRG HEVLIQAGAG EGSAITDADF KAAGAQLVGT
ADQVWADADL LLKVKEPIAA EYGRLRHGQI LFTFLHLAAS RACTDALLDS GTTSIAYETV
QTADGALPLL APMSEVAGRL AAQVGAYHLM RTQGGRGVLM GGVPGVEPAD VVVIGAGTAG
YNAARIANGM GATVTVLDIN IDKLRQLDAE FCGRIHTRYS SAYELEGAVK RADLVIGAVL
VPGAKAPKLV SNSLVAHMKP GAVLVDIAID QGGCFEGSRP TTYDHPTFAV HDTLFYCVAN
MPASVPKTST YALTNATMPY VLELADHGWR AACRSNPALA KGLSTHEGAL LSERVATDLG
VPFTEPASVL A
