NANA_STAA8
ID NANA_STAA8 Reviewed; 293 AA.
AC Q2G160;
DT 05-FEB-2008, integrated into UniProtKB/Swiss-Prot.
DT 21-MAR-2006, sequence version 1.
DT 03-AUG-2022, entry version 97.
DE RecName: Full=N-acetylneuraminate lyase {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:23418011};
DE Short=NAL {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:23418011};
DE Short=Neu5Ac lyase {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:23418011};
DE EC=4.1.3.3 {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000269|PubMed:23418011};
DE AltName: Full=N-acetylneuraminate pyruvate-lyase {ECO:0000255|HAMAP-Rule:MF_01237};
DE AltName: Full=N-acetylneuraminic acid aldolase {ECO:0000255|HAMAP-Rule:MF_01237};
DE AltName: Full=Sialate lyase {ECO:0000255|HAMAP-Rule:MF_01237};
DE AltName: Full=Sialic acid aldolase {ECO:0000255|HAMAP-Rule:MF_01237};
DE AltName: Full=Sialic acid lyase {ECO:0000255|HAMAP-Rule:MF_01237};
GN Name=nanA {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:23418011};
GN OrderedLocusNames=SAOUHSC_00295;
OS Staphylococcus aureus (strain NCTC 8325 / PS 47).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Staphylococcaceae;
OC Staphylococcus.
OX NCBI_TaxID=93061;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=NCTC 8325 / PS 47;
RA Gillaspy A.F., Worrell V., Orvis J., Roe B.A., Dyer D.W., Iandolo J.J.;
RT "The Staphylococcus aureus NCTC 8325 genome.";
RL (In) Fischetti V., Novick R., Ferretti J., Portnoy D., Rood J. (eds.);
RL Gram positive pathogens, 2nd edition, pp.381-412, ASM Press, Washington
RL D.C. (2006).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF APOENZYME, WILD-TYPE AND MUTANTS
RP CYS-165 AND GAMMA-THIALYSINE-165 IN COMPLEX WITH PYRUVATE, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF LYS-165,
RP AND ACTIVE SITE.
RC STRAIN=NCTC 8325 / PS 47;
RX PubMed=23418011; DOI=10.1002/cbic.201200714;
RA Timms N., Windle C.L., Polyakova A., Ault J.R., Trinh C.H., Pearson A.R.,
RA Nelson A., Berry A.;
RT "Structural insights into the recovery of aldolase activity in N-
RT acetylneuraminic acid lyase by replacement of the catalytically active
RT lysine with gamma-thialysine by using a chemical mutagenesis strategy.";
RL ChemBioChem 14:474-481(2013).
CC -!- FUNCTION: Catalyzes the reversible aldol cleavage of N-acetylneuraminic
CC acid (sialic acid; Neu5Ac) to form pyruvate and N-acetylmannosamine
CC (ManNAc) via a Schiff base intermediate. {ECO:0000255|HAMAP-
CC Rule:MF_01237, ECO:0000269|PubMed:23418011}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aceneuramate = N-acetyl-D-mannosamine + pyruvate;
CC Xref=Rhea:RHEA:23296, ChEBI:CHEBI:15361, ChEBI:CHEBI:17122,
CC ChEBI:CHEBI:173083; EC=4.1.3.3; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_01237, ECO:0000269|PubMed:23418011};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.2 mM for N-acetylneuraminate (at pH 7.4)
CC {ECO:0000269|PubMed:23418011};
CC KM=2.4 mM for N-acetylneuraminate (at pH 6.8)
CC {ECO:0000269|PubMed:23418011};
CC Note=kcat is 250 min(-1) for the cleavage of Neu5Ac at pH 7.4, and is
CC 260 min(-1) at pH 6.8. {ECO:0000269|PubMed:23418011};
CC pH dependence:
CC Optimum pH is 7.4. {ECO:0000269|PubMed:23418011};
CC -!- PATHWAY: Amino-sugar metabolism; N-acetylneuraminate degradation; D-
CC fructose 6-phosphate from N-acetylneuraminate: step 1/5.
CC {ECO:0000255|HAMAP-Rule:MF_01237}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000255|HAMAP-Rule:MF_01237,
CC ECO:0000269|PubMed:23418011}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_01237}.
CC -!- MISCELLANEOUS: When the catalytic Lys-165 residue is replaced by the
CC unnatural amino acid gamma-thialysine, the enzyme regains significant
CC activity compared with the Cys-165 mutant; regains of approximately 17%
CC of the wild-type catalytic efficiency. {ECO:0000269|PubMed:23418011}.
CC -!- SIMILARITY: Belongs to the DapA family. NanA subfamily.
CC {ECO:0000255|HAMAP-Rule:MF_01237}.
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DR EMBL; CP000253; ABD29464.1; -; Genomic_DNA.
DR RefSeq; WP_001030738.1; NZ_LS483365.1.
DR RefSeq; YP_498885.1; NC_007795.1.
DR PDB; 4AH7; X-ray; 2.30 A; A/B/C/D=2-293.
DR PDB; 4AHO; X-ray; 2.00 A; A/B/C/D=1-293.
DR PDB; 4AHP; X-ray; 2.10 A; A/B/C/D=2-293.
DR PDB; 4AHQ; X-ray; 1.95 A; A/B/C/D=2-293.
DR PDB; 4AMA; X-ray; 2.35 A; A/B/C/D=2-293.
DR PDB; 5A8G; X-ray; 1.72 A; A/B=2-293.
DR PDB; 5KZD; X-ray; 2.33 A; A/B/C/D=1-293.
DR PDB; 5KZE; X-ray; 1.74 A; A/B/C/D/E/F/G/H=1-293.
DR PDB; 5LKY; X-ray; 1.70 A; A/B/C/D=2-293.
DR PDBsum; 4AH7; -.
DR PDBsum; 4AHO; -.
DR PDBsum; 4AHP; -.
DR PDBsum; 4AHQ; -.
DR PDBsum; 4AMA; -.
DR PDBsum; 5A8G; -.
DR PDBsum; 5KZD; -.
DR PDBsum; 5KZE; -.
DR PDBsum; 5LKY; -.
DR AlphaFoldDB; Q2G160; -.
DR SMR; Q2G160; -.
DR STRING; 1280.SAXN108_0299; -.
DR EnsemblBacteria; ABD29464; ABD29464; SAOUHSC_00295.
DR GeneID; 3918976; -.
DR KEGG; sao:SAOUHSC_00295; -.
DR PATRIC; fig|93061.5.peg.268; -.
DR eggNOG; COG0329; Bacteria.
DR HOGENOM; CLU_049343_5_1_9; -.
DR OMA; VVPPVCT; -.
DR BRENDA; 4.1.3.3; 3352.
DR UniPathway; UPA00629; UER00680.
DR PRO; PR:Q2G160; -.
DR Proteomes; UP000008816; Chromosome.
DR GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR GO; GO:0008747; F:N-acetylneuraminate lyase activity; IBA:GO_Central.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-UniRule.
DR GO; GO:0019262; P:N-acetylneuraminate catabolic process; IBA:GO_Central.
DR CDD; cd00954; NAL; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR HAMAP; MF_01237; N_acetylneuram_lyase; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR002220; DapA-like.
DR InterPro; IPR005264; NanA.
DR InterPro; IPR020625; Schiff_base-form_aldolases_AS.
DR Pfam; PF00701; DHDPS; 1.
DR PIRSF; PIRSF001365; DHDPS; 1.
DR PRINTS; PR00146; DHPICSNTHASE.
DR SMART; SM01130; DHDPS; 1.
DR PROSITE; PS00666; DHDPS_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cytoplasm; Lyase;
KW Reference proteome; Schiff base.
FT CHAIN 1..293
FT /note="N-acetylneuraminate lyase"
FT /id="PRO_1000066938"
FT ACT_SITE 165
FT /note="Schiff-base intermediate with substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01237,
FT ECO:0000269|PubMed:23418011"
FT BINDING 48..49
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01237,
FT ECO:0000269|PubMed:23418011"
FT SITE 137
FT /note="Involved in proton transfer during cleavage"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01237"
FT MUTAGEN 165
FT /note="K->C: 3125-fold decrease in catalytic activity, and
FT 3-fold increase in substrate affinity."
FT /evidence="ECO:0000269|PubMed:23418011"
FT STRAND 7..11
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 24..36
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 41..47
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 48..50
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 52..54
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 57..70
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 75..80
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 86..99
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 102..107
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 116..130
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 134..138
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 140..143
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 149..156
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 161..166
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 171..180
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 184..188
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 191..193
FT /evidence="ECO:0007829|PDB:5A8G"
FT HELIX 194..199
FT /evidence="ECO:0007829|PDB:5LKY"
FT STRAND 203..208
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 210..225
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 229..249
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 251..261
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 278..280
FT /evidence="ECO:0007829|PDB:5LKY"
FT HELIX 281..291
FT /evidence="ECO:0007829|PDB:5LKY"
SQ SEQUENCE 293 AA; 33043 MW; 62D7DEFFDC1AF431 CRC64;
MNKDLKGLYA ALLVPFDENG QVNEQGLKQI AQNAIETEEL DGLYVNGSSG ENFLLNTEQK
KQVFKVAKEA VGDKVKLIAQ VGSLDLNEAI ELGKYATELG YDALSAVTPF YYPFTFEEIR
DYYFDIIEAT QNNMIIYAIP DLTGVNISIE QFSELFNHEK IVGVKYTAPN FFLLERIRKA
FPDKLILSGF DEMLVQATIS GVDGAIGSTY NVNGRRARKI FDLARQGQIQ EAYQLQHDSN
DIIETVLSMG IYPTLKEILR HRGIDAGLPK RPFKPFNEAH RQTLDQLIAK YDL
