ID NANA_ECOLI Reviewed; 297 AA.
AC P0A6L4; P06995; Q2M8Y8;
DT 01-APR-1988, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 27-MAR-2024, entry version 142.
DE RecName: Full=N-acetylneuraminate lyase {ECO:0000303|PubMed:3909108};
DE Short=AcNeu lyase {ECO:0000303|PubMed:1646603};
DE Short=NAL {ECO:0000303|PubMed:9047371};
DE Short=Neu5Ac lyase {ECO:0000303|PubMed:8081752};
DE EC=4.1.3.3 {ECO:0000269|PubMed:12711733, ECO:0000269|PubMed:1646603, ECO:0000269|PubMed:24521460, ECO:0000269|PubMed:33895133};
DE AltName: Full=N-acetylneuraminate pyruvate-lyase {ECO:0000303|PubMed:3909108};
DE AltName: Full=N-acetylneuraminic acid aldolase {ECO:0000303|PubMed:3909108};
DE AltName: Full=NALase {ECO:0000303|Ref.2};
DE AltName: Full=Sialate lyase;
DE AltName: Full=Sialic acid aldolase;
DE AltName: Full=Sialic acid lyase;
GN Name=nanA {ECO:0000303|Ref.2}; Synonyms=npl {ECO:0000303|PubMed:3909108};
GN OrderedLocusNames=b3225, JW3194;
OS Escherichia coli (strain K12).
OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PARTIAL PROTEIN SEQUENCE.
RC STRAIN=K12;
RX PubMed=3909108; DOI=10.1093/nar/13.24.8843;
RA Ohta Y., Watanabe K., Kimura A.;
RT "Complete nucleotide sequence of the E. coli N-acetylneuraminate lyase.";
RL Nucleic Acids Res. 13:8843-8852(1985).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=JE1011;
RA Kawakami B., Kudo T., Narahashi Y., Horikoshi K.;
RT "Nucleotide sequence of the N-acetylneuraminate lyase gene of Escherichia
RT coli.";
RL Agric. Biol. Chem. 50:2155-2158(1986).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [5]
RP PROTEIN SEQUENCE OF 2-24, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=K12 / C600 / SF8;
RX PubMed=1646603; DOI=10.1042/bj2760541;
RA Aisaka K., Igarashi A., Yamaguchi K., Uwajima T.;
RT "Purification, crystallization and characterization of N-acetylneuraminate
RT lyase from Escherichia coli.";
RL Biochem. J. 276:541-546(1991).
RN [6]
RP IDENTIFICATION BY 2D-GEL.
RX PubMed=9298644; DOI=10.1002/elps.1150180805;
RA VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.;
RT "Escherichia coli proteome analysis using the gene-protein database.";
RL Electrophoresis 18:1243-1251(1997).
RN [7]
RP INDUCTION.
RX PubMed=23935044; DOI=10.1128/jb.00692-13;
RA Kalivoda K.A., Steenbergen S.M., Vimr E.R.;
RT "Control of the Escherichia coli sialoregulon by transcriptional repressor
RT NanR.";
RL J. Bacteriol. 195:4689-4701(2013).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND ACTIVITY REGULATION.
RC STRAIN=K12;
RX PubMed=33895133; DOI=10.1016/j.jbc.2021.100699;
RA Kentache T., Thabault L., Deumer G., Haufroid V., Frederick R.,
RA Linster C.L., Peracchi A., Veiga-da-Cunha M., Bommer G.T.,
RA Van Schaftingen E.;
RT "The metalloprotein YhcH is an anomerase providing N-acetylneuraminate
RT aldolase with the open form of its substrate.";
RL J. Biol. Chem. 296:100699-100699(2021).
RN [9] {ECO:0007744|PDB:1NAL}
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS), SUBUNIT, AND ACTIVE SITE.
RX PubMed=8081752; DOI=10.1016/s0969-2126(00)00038-1;
RA Izard T., Lawrence M.C., Malby R.L., Lilley G.G., Colman P.M.;
RT "The three-dimensional structure of N-acetylneuraminate lyase from
RT Escherichia coli.";
RL Structure 2:361-369(1994).
RN [10] {ECO:0007744|PDB:1FDY, ECO:0007744|PDB:1FDZ}
RP X-RAY CRYSTALLOGRAPHY (2.45 ANGSTROMS) OF COMPLEXES WITH HYDROXYPYRUVATE
RP AND PYRUVATE, ACTIVITY REGULATION, SUBUNIT, AND ACTIVE SITE.
RX PubMed=9047371; DOI=10.1006/jmbi.1996.0769;
RA Lawrence M.C., Barbosa J.A.R.G., Smith B.J., Hall N.E., Pilling P.A.,
RA Ooi H.C., Marcuccio S.M.;
RT "Structure and mechanism of a sub-family of enzymes related to N-
RT acetylneuraminate lyase.";
RL J. Mol. Biol. 266:381-399(1997).
RN [11] {ECO:0007744|PDB:1HL2}
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF MUTANT ARG-142 IN COMPLEX WITH
RP 3-HYDROXYPYRUVATE, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, MUTAGENESIS OF LEU-142, AND ACTIVE
RP SITE.
RX PubMed=12711733; DOI=10.1073/pnas.0531477100;
RA Joerger A.C., Mayer S., Fersht A.R.;
RT "Mimicking natural evolution in vitro: an N-acetylneuraminate lyase mutant
RT with an increased dihydrodipicolinate synthase activity.";
RL Proc. Natl. Acad. Sci. U.S.A. 100:5694-5699(2003).
RN [12] {ECO:0007744|PDB:2WKJ}
RP X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) OF 2-296 OF MUTANT ASN-192 IN
RP COMPLEX WITH PYRUVATE, SUBUNIT, ACTIVE SITE, AND MUTAGENESIS OF GLU-192.
RX PubMed=19923724; DOI=10.1107/s1744309109037403;
RA Campeotto I., Carr S.B., Trinh C.H., Nelson A.S., Berry A., Phillips S.E.,
RA Pearson A.R.;
RT "Structure of an Escherichia coli N-acetyl-D-neuraminic acid lyase mutant,
RT E192N, in complex with pyruvate at 1.45 angstrom resolution.";
RL Acta Crystallogr. F Struct. Biol. Commun. 65:1088-1090(2009).
RN [13] {ECO:0007744|PDB:4BWL}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 2-297 OF MUTANT ALA-137 IN
RP COMPLEX WITH N-ACETYLNEURAMINIC ACID AND N-ACETYL-D-MANNOSAMINE, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, ACTIVE SITE,
RP AND MUTAGENESIS OF SER-47; THR-48; TYR-110; TYR-137; THR-167 AND PHE-252.
RX PubMed=24521460; DOI=10.1021/cb500067z;
RA Daniels A.D., Campeotto I., van der Kamp M.W., Bolt A.H., Trinh C.H.,
RA Phillips S.E., Pearson A.R., Nelson A., Mulholland A.J., Berry A.;
RT "Reaction mechanism of N-acetylneuraminic acid lyase revealed by a
RT combination of crystallography, QM/MM simulation, and mutagenesis.";
RL ACS Chem. Biol. 9:1025-1032(2014).
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 (PubMed:1646603,
CC PubMed:33895133, PubMed:12711733, PubMed:24521460). Experiments show
CC the true substrate is aceneuramate (linearized Neu5Ac), which forms
CC spontaneously at alkaline pH (PubMed:33895133). Linear aceneuramate can
CC be provided by NanQ (PubMed:33895133). Can also cleave other substrates
CC such as N-glycollylneuraminic acid (GcNeu), but not colominic acid or
CC 2-oxocarboxylic acids such as 2-oxohexanoic acid, 2-oxo-octanoic acid,
CC 2-oxo-3-deoxyoctanoic acid and 2-oxononanoic acid (PubMed:1646603).
CC {ECO:0000269|PubMed:12711733, ECO:0000269|PubMed:1646603,
CC ECO:0000269|PubMed:24521460, ECO:0000269|PubMed:33895133}.
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;
CC Evidence={ECO:0000269|PubMed:12711733, ECO:0000269|PubMed:1646603,
CC ECO:0000269|PubMed:24521460, ECO:0000269|PubMed:33895133};
CC -!- ACTIVITY REGULATION: Inhibited by reduction with NaBH(4), and by Cu(2+)
CC ions, p-chloromercuribenzoate and N-bromosuccinimide (PubMed:1646603).
CC Inhibited by beta-hydroxypyruvate (PubMed:9047371, PubMed:12711733).
CC Co(2+), Mn(2+) and Ni(2+) stimulate activity, perhaps by enhancing the
CC opening of the substrate Neu5Ac (PubMed:33895133).
CC {ECO:0000269|PubMed:12711733, ECO:0000269|PubMed:1646603,
CC ECO:0000269|PubMed:33895133, ECO:0000269|PubMed:9047371}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.3 mM for N-acetylneuraminate {ECO:0000269|PubMed:1646603};
CC KM=2.5 mM for N-acetylneuraminate {ECO:0000269|PubMed:12711733};
CC KM=2.2 mM for N-acetylneuraminate {ECO:0000269|PubMed:24521460};
CC KM=3.3 mM for N-glycollylneuraminate {ECO:0000269|PubMed:1646603};
CC Vmax=71.4 umol/min/mg enzyme with N-acetylneuraminate as substrate
CC {ECO:0000269|PubMed:1646603};
CC Vmax=14.0 umol/min/mg enzyme with N-glycollylneuraminate as substrate
CC {ECO:0000269|PubMed:1646603};
CC Note=kcat is 7.7 sec(-1) with N-acetylneuraminate as substrate
CC (PubMed:12711733). kcat is 0.47 sec(-1) for the aldol condensation of
CC L-aspartate beta-semialdehyde and pyruvate (at pH 7.0)
CC (PubMed:12711733). kcat is 0.27 sec(-1) for the aldol condensation of
CC L-aspartate beta-semialdehyde and pyruvate (at pH 8.0)
CC (PubMed:12711733). kcat is 250 min(-1) with N-acetylneuraminate as
CC substrate (PubMed:24521460). {ECO:0000269|PubMed:12711733,
CC ECO:0000269|PubMed:24521460};
CC pH dependence:
CC Optimum pH is 6.5-7.0. {ECO:0000269|PubMed:1646603};
CC Temperature dependence:
CC Optimum temperature is 80 degrees Celsius.
CC {ECO:0000269|PubMed:1646603};
CC -!- PATHWAY: Amino-sugar metabolism; N-acetylneuraminate degradation; D-
CC fructose 6-phosphate from N-acetylneuraminate: step 1/5. {ECO:0000305}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:12711733,
CC ECO:0000269|PubMed:19923724, ECO:0000269|PubMed:24521460,
CC ECO:0000269|PubMed:8081752, ECO:0000269|PubMed:9047371}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- INDUCTION: Negatively regulated by the transcriptional repressor NanR.
CC Induced by N-acetylneuraminate, via inactivation of NanR.
CC {ECO:0000269|PubMed:23935044}.
CC -!- SIMILARITY: Belongs to the DapA family. NanA subfamily.
CC {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000305}.
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DR EMBL; X03345; CAA27051.1; -; Genomic_DNA.
DR EMBL; D00067; BAA00046.1; -; Genomic_DNA.
DR EMBL; U18997; AAA58027.1; -; Genomic_DNA.
DR EMBL; U00096; AAC76257.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE77268.1; -; Genomic_DNA.
DR PIR; JP0002; WZECN.
DR RefSeq; NP_417692.1; NC_000913.3.
DR RefSeq; WP_000224714.1; NZ_STEB01000012.1.
DR PDB; 1FDY; X-ray; 2.45 A; A/B/C/D=1-297.
DR PDB; 1FDZ; X-ray; 2.60 A; A/B/C/D=1-297.
DR PDB; 1HL2; X-ray; 1.80 A; A/B/C/D=1-297.
DR PDB; 1NAL; X-ray; 2.20 A; 1/2/3/4=1-297.
DR PDB; 2WKJ; X-ray; 1.45 A; A/B/C/D=2-296.
DR PDB; 2WNN; X-ray; 1.65 A; A/B/C/D=2-296.
DR PDB; 2WNQ; X-ray; 1.80 A; A/B/C/D=2-297.
DR PDB; 2WNZ; X-ray; 1.85 A; A/B/C/D=2-297.
DR PDB; 2WO5; X-ray; 2.20 A; A/B/C/D=2-297.
DR PDB; 2WPB; X-ray; 2.05 A; A/B/C/D=2-297.
DR PDB; 2XFW; X-ray; 1.65 A; A/B/C/D=2-297.
DR PDB; 2YGY; X-ray; 1.90 A; A/B/C/D=2-297.
DR PDB; 3LBC; X-ray; 1.85 A; A/B/C/D=1-297.
DR PDB; 3LBM; X-ray; 1.48 A; A/B/C/D=1-297.
DR PDB; 3LCF; X-ray; 1.86 A; A/B/C/D=1-297.
DR PDB; 3LCG; X-ray; 1.78 A; A/B/C/D=1-297.
DR PDB; 3LCH; X-ray; 2.04 A; A/B/C/D=1-297.
DR PDB; 3LCI; X-ray; 2.12 A; A/B/C/D=1-297.
DR PDB; 3LCL; X-ray; 1.83 A; A/B/C/D=1-297.
DR PDB; 3LCW; X-ray; 2.35 A; A/B/C/D=1-297.
DR PDB; 3LCX; X-ray; 1.98 A; A/B/C/D=1-297.
DR PDB; 4BWL; X-ray; 2.00 A; A/B/C/D=2-297.
DR PDB; 4UUI; X-ray; 1.79 A; A/B/C/D=2-297.
DR PDBsum; 1FDY; -.
DR PDBsum; 1FDZ; -.
DR PDBsum; 1HL2; -.
DR PDBsum; 1NAL; -.
DR PDBsum; 2WKJ; -.
DR PDBsum; 2WNN; -.
DR PDBsum; 2WNQ; -.
DR PDBsum; 2WNZ; -.
DR PDBsum; 2WO5; -.
DR PDBsum; 2WPB; -.
DR PDBsum; 2XFW; -.
DR PDBsum; 2YGY; -.
DR PDBsum; 3LBC; -.
DR PDBsum; 3LBM; -.
DR PDBsum; 3LCF; -.
DR PDBsum; 3LCG; -.
DR PDBsum; 3LCH; -.
DR PDBsum; 3LCI; -.
DR PDBsum; 3LCL; -.
DR PDBsum; 3LCW; -.
DR PDBsum; 3LCX; -.
DR PDBsum; 4BWL; -.
DR PDBsum; 4UUI; -.
DR AlphaFoldDB; P0A6L4; -.
DR SMR; P0A6L4; -.
DR BioGRID; 4262442; 344.
DR DIP; DIP-10302N; -.
DR IntAct; P0A6L4; 5.
DR STRING; 511145.b3225; -.
DR jPOST; P0A6L4; -.
DR PaxDb; 511145-b3225; -.
DR EnsemblBacteria; AAC76257; AAC76257; b3225.
DR GeneID; 83578187; -.
DR GeneID; 947742; -.
DR KEGG; ecj:JW3194; -.
DR KEGG; eco:b3225; -.
DR PATRIC; fig|1411691.4.peg.3503; -.
DR EchoBASE; EB0631; -.
DR eggNOG; COG0329; Bacteria.
DR HOGENOM; CLU_049343_6_0_6; -.
DR InParanoid; P0A6L4; -.
DR OMA; TGEFTTM; -.
DR OrthoDB; 199953at2; -.
DR PhylomeDB; P0A6L4; -.
DR BioCyc; EcoCyc:ACNEULY-MONOMER; -.
DR BioCyc; MetaCyc:ACNEULY-MONOMER; -.
DR BRENDA; 4.1.3.3; 2026.
DR SABIO-RK; P0A6L4; -.
DR UniPathway; UPA00629; UER00680.
DR EvolutionaryTrace; P0A6L4; -.
DR PRO; PR:P0A6L4; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR GO; GO:0008747; F:N-acetylneuraminate lyase activity; IDA:EcoCyc.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-UniRule.
DR GO; GO:0019262; P:N-acetylneuraminate catabolic process; IMP:EcoCyc.
DR GO; GO:0044010; P:single-species biofilm formation; IMP:EcoCyc.
DR CDD; cd00954; NAL; 1.
DR Gene3D; 3.20.20.70; Aldolase class I; 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 InterPro; IPR020624; Schiff_base-form_aldolases_CS.
DR NCBIfam; TIGR00683; nanA; 1.
DR PANTHER; PTHR42849; N-ACETYLNEURAMINATE LYASE; 1.
DR PANTHER; PTHR42849:SF1; N-ACETYLNEURAMINATE LYASE; 1.
DR Pfam; PF00701; DHDPS; 1.
DR PIRSF; PIRSF001365; DHDPS; 1.
DR PRINTS; PR00146; DHPICSNTHASE.
DR SMART; SM01130; DHDPS; 1.
DR SUPFAM; SSF51569; Aldolase; 1.
DR PROSITE; PS00665; DHDPS_1; 1.
DR PROSITE; PS00666; DHDPS_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cytoplasm;
KW Direct protein sequencing; Lyase; Reference proteome; Schiff base.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:1646603"
FT CHAIN 2..297
FT /note="N-acetylneuraminate lyase"
FT /id="PRO_0000103209"
FT ACT_SITE 137
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:24521460"
FT ACT_SITE 165
FT /note="Schiff-base intermediate with substrate"
FT /evidence="ECO:0000269|PubMed:12711733,
FT ECO:0000269|PubMed:19923724, ECO:0000269|PubMed:24521460,
FT ECO:0000269|PubMed:9047371, ECO:0000305|PubMed:8081752"
FT BINDING 47
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 47
FT /ligand="pyruvate"
FT /ligand_id="ChEBI:CHEBI:15361"
FT /evidence="ECO:0000269|PubMed:9047371,
FT ECO:0000305|PubMed:12711733, ECO:0007744|PDB:1FDY,
FT ECO:0007744|PDB:1FDZ, ECO:0007744|PDB:1HL2"
FT BINDING 48
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 48
FT /ligand="pyruvate"
FT /ligand_id="ChEBI:CHEBI:15361"
FT /evidence="ECO:0000269|PubMed:9047371,
FT ECO:0000305|PubMed:12711733, ECO:0007744|PDB:1FDY,
FT ECO:0007744|PDB:1FDZ, ECO:0007744|PDB:1HL2"
FT BINDING 167
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 167
FT /ligand="N-acetyl-D-mannosamine"
FT /ligand_id="ChEBI:CHEBI:17122"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 189
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 189
FT /ligand="N-acetyl-D-mannosamine"
FT /ligand_id="ChEBI:CHEBI:17122"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 191
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 191
FT /ligand="N-acetyl-D-mannosamine"
FT /ligand_id="ChEBI:CHEBI:17122"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 192
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 192
FT /ligand="N-acetyl-D-mannosamine"
FT /ligand_id="ChEBI:CHEBI:17122"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 208
FT /ligand="aceneuramate"
FT /ligand_id="ChEBI:CHEBI:173083"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT BINDING 208
FT /ligand="N-acetyl-D-mannosamine"
FT /ligand_id="ChEBI:CHEBI:17122"
FT /evidence="ECO:0000269|PubMed:24521460,
FT ECO:0007744|PDB:4BWL"
FT SITE 47
FT /note="Required to correctly position the proton donor"
FT /evidence="ECO:0000305|PubMed:24521460"
FT SITE 110
FT /note="Required to correctly position the proton donor"
FT /evidence="ECO:0000305|PubMed:24521460"
FT MUTAGEN 47
FT /note="S->A: 21-fold decrease in catalytic efficiency for
FT the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 47
FT /note="S->C: 40-fold decrease in catalytic efficiency for
FT the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 47
FT /note="S->T: No significant change in kinetic parameters
FT for the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 48
FT /note="T->A,S: Slight increase in catalytic efficiency for
FT the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 110
FT /note="Y->A: 40-fold decrease in catalytic efficiency for
FT the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 110
FT /note="Y->F: No significant change in kinetic parameters
FT for the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 137
FT /note="Y->A: Loss of Neu5Ac cleavage activity. Is still
FT able to form a Schiff base with the substrate."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 137
FT /note="Y->F: Retains very low Neu5Ac cleavage activity."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 142
FT /note="L->R: Changes substrate preference. Maintains much
FT of its original N-acetylneuraminate lyase activity, but
FT shows a 19-fold increase in condensation of L-aspartate
FT beta-semialdehyde (L-ASA) and pyruvate."
FT /evidence="ECO:0000269|PubMed:12711733"
FT MUTAGEN 167
FT /note="T->A: 4-fold decrease in catalytic efficiency for
FT the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 167
FT /note="T->S: No significant change in kinetic parameters
FT for the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT MUTAGEN 192
FT /note="E->N: 6-fold higher specificity for
FT dipropylaminocarbonyl-substituted derivatives."
FT /evidence="ECO:0000269|PubMed:19923724"
FT MUTAGEN 252
FT /note="F->A,Y: No significant change in kinetic parameters
FT for the cleavage of Neu5Ac."
FT /evidence="ECO:0000269|PubMed:24521460"
FT CONFLICT 70
FT /note="A -> G (in Ref. 1; CAA27051)"
FT /evidence="ECO:0000305"
FT CONFLICT 84
FT /note="S -> T (in Ref. 1; CAA27051)"
FT /evidence="ECO:0000305"
FT CONFLICT 282
FT /note="L -> Q (in Ref. 1; CAA27051)"
FT /evidence="ECO:0000305"
FT HELIX 2..5
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 7..11
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 20..22
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 24..36
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 40..46
FT /evidence="ECO:0007829|PDB:2WKJ"
FT TURN 47..50
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 51..53
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 56..70
FT /evidence="ECO:0007829|PDB:2WKJ"
FT TURN 71..73
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 74..79
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 85..98
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 101..106
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 115..129
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 134..138
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 140..143
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 149..156
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 161..166
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 171..180
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 185..188
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 191..193
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 194..200
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 204..207
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 210..226
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 229..249
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 251..261
FT /evidence="ECO:0007829|PDB:2WKJ"
FT STRAND 265..267
FT /evidence="ECO:0007829|PDB:4BWL"
FT HELIX 279..281
FT /evidence="ECO:0007829|PDB:2WKJ"
FT HELIX 282..295
FT /evidence="ECO:0007829|PDB:2WKJ"
SQ SEQUENCE 297 AA; 32593 MW; BA9F30B4A7624167 CRC64;
MATNLRGVMA ALLTPFDQQQ ALDKASLRRL VQFNIQQGID GLYVGGSTGE AFVQSLSERE
QVLEIVAEEA KGKIKLIAHV GCVSTAESQQ LAASAKRYGF DAVSAVTPFY YPFSFEEHCD
HYRAIIDSAD GLPMVVYNIP ALSGVKLTLD QINTLVTLPG VGALKQTSGD LYQMEQIRRE
HPDLVLYNGY DEIFASGLLA GADGGIGSTY NIMGWRYQGI VKALKEGDIQ TAQKLQTECN
KVIDLLIKTG VFRGLKTVLH YMDVVSVPLC RKPFGPVDEK YLPELKALAQ QLMQERG
//
