PL_STRMK
ID PL_STRMK Reviewed; 331 AA.
AC B2FHL8;
DT 09-JUL-2014, integrated into UniProtKB/Swiss-Prot.
DT 10-JUN-2008, sequence version 1.
DT 25-MAY-2022, entry version 66.
DE RecName: Full=Polysaccharide lyase {ECO:0000303|PubMed:24257754};
DE Short=PL {ECO:0000303|PubMed:24257754};
DE EC=4.2.2.- {ECO:0000269|PubMed:24257754};
DE AltName: Full=Alginate lyase {ECO:0000305|PubMed:24257754};
DE EC=4.2.2.3 {ECO:0000269|PubMed:24257754};
DE AltName: Full=Endolytic polysaccharide lyase {ECO:0000305};
DE AltName: Full=Hyaluronate lyase {ECO:0000305|PubMed:24257754};
DE EC=4.2.2.1 {ECO:0000269|PubMed:24257754};
DE AltName: Full=Multifunctional polysaccharide lyase {ECO:0000303|PubMed:24808176};
DE AltName: Full=Poly-beta-D-glucuronate lyase {ECO:0000305|PubMed:24257754};
DE EC=4.2.2.14 {ECO:0000269|PubMed:24257754};
DE Flags: Precursor;
GN OrderedLocusNames=Smlt1473;
OS Stenotrophomonas maltophilia (strain K279a).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales;
OC Xanthomonadaceae; Stenotrophomonas; Stenotrophomonas maltophilia group.
OX NCBI_TaxID=522373;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K279a;
RX PubMed=18419807; DOI=10.1186/gb-2008-9-4-r74;
RA Crossman L.C., Gould V.C., Dow J.M., Vernikos G.S., Okazaki A.,
RA Sebaihia M., Saunders D., Arrowsmith C., Carver T., Peters N., Adlem E.,
RA Kerhornou A., Lord A., Murphy L., Seeger K., Squares R., Rutter S.,
RA Quail M.A., Rajandream M.A., Harris D., Churcher C., Bentley S.D.,
RA Parkhill J., Thomson N.R., Avison M.B.;
RT "The complete genome, comparative and functional analysis of
RT Stenotrophomonas maltophilia reveals an organism heavily shielded by drug
RT resistance determinants.";
RL Genome Biol. 9:R74.1-R74.13(2008).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, SUBCELLULAR LOCATION, AND MUTAGENESIS OF
RP CYS-23; ASN-167; HIS-168; ARG-215 AND TYR-222.
RC STRAIN=K279a;
RX PubMed=24257754; DOI=10.1074/jbc.m113.489195;
RA MacDonald L.C., Berger B.W.;
RT "A polysaccharide lyase from Stenotrophomonas maltophilia with a unique,
RT pH-regulated substrate specificity.";
RL J. Biol. Chem. 289:312-325(2014).
RN [3]
RP 3D-STRUCTURE MODELING, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF
RP TRP-171; HIS-221 AND ARG-312.
RC STRAIN=K279a;
RX PubMed=24808176; DOI=10.1074/jbc.m114.571299;
RA MacDonald L.C., Berger B.W.;
RT "Insight into the role of substrate-binding residues in conferring
RT substrate specificity for the multifunctional polysaccharide lyase
RT Smlt1473.";
RL J. Biol. Chem. 289:18022-18032(2014).
CC -!- FUNCTION: Polysaccharide lyase that catalyzes the depolymerization of
CC several anionic polysaccharides via a beta-elimination mechanism.
CC Exhibits broad substrate specificity, catalyzing the degradation of not
CC only alginate and poly-beta-D-mannuronate (poly-ManA), but poly-beta-D-
CC glucuronate (poly-GlcA or poly-GlcUA) and hyaluronate (HA) as well. The
CC oligosaccharide products formed by enzymatic cleavage are comprised
CC mainly of disaccharides, with a lower abundance of trimers and
CC pentamers. Is not active on poly-D-galacturonate, heparin and heparin
CC sulfate. {ECO:0000269|PubMed:24257754}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Eliminative cleavage of alginate to give oligosaccharides with
CC 4-deoxy-alpha-L-erythro-hex-4-enuronosyl groups at their non-reducing
CC ends and beta-D-mannuronate at their reducing end.; EC=4.2.2.3;
CC Evidence={ECO:0000269|PubMed:24257754};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[hyaluronan](n) = n 3-(4-deoxy-beta-D-gluc-4-enuronosyl)-N-
CC acetyl-D-glucosamine + H2O; Xref=Rhea:RHEA:50240, Rhea:RHEA-
CC COMP:12583, ChEBI:CHEBI:15377, ChEBI:CHEBI:132151,
CC ChEBI:CHEBI:132153; EC=4.2.2.1;
CC Evidence={ECO:0000269|PubMed:24257754};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Eliminative cleavage of (1->4)-beta-D-glucuronans to give
CC oligosaccharides with 4-deoxy-beta-D-gluc-4-enuronosyl groups at
CC their non-reducing ends. Complete degradation of glucuronans results
CC in the formation of tetrasaccharides.; EC=4.2.2.14;
CC Evidence={ECO:0000269|PubMed:24257754};
CC -!- ACTIVITY REGULATION: Is inhibited by mono- and divalent cations as well
CC as L-ascorbic acid 6-hexadecanoate. {ECO:0000269|PubMed:24257754}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.14 mg/ml for poly-GlcA {ECO:0000269|PubMed:24257754};
CC KM=0.26 mg/ml for poly-ManA {ECO:0000269|PubMed:24257754};
CC KM=0.55 mg/ml for hyaluronan {ECO:0000269|PubMed:24257754};
CC KM=0.17 mM for poly-GlcA (at pH 7) {ECO:0000269|PubMed:24808176};
CC KM=0.35 mM for poly-ManA (at pH 9) {ECO:0000269|PubMed:24808176};
CC Note=Vmax for poly-GlcA is about 10-fold greater versus poly-ManA or
CC HA (PubMed:24257754). kcat is 31.9 sec(-1) with poly-GlcA as
CC substrate (at pH 7) (PubMed:24808176). kcat is 3.3 sec(-1) with poly-
CC ManA as substrate (at pH 9) (PubMed:24808176).
CC {ECO:0000269|PubMed:24257754, ECO:0000269|PubMed:24808176};
CC pH dependence:
CC Optimum pH for enzymatic activity is substrate-dependent, with
CC optimal hyaluronate degradation at pH 5, poly-beta-D-glucuronate
CC degradation at pH 7, and alginate degradation at pH 9.
CC {ECO:0000269|PubMed:24257754};
CC -!- SUBCELLULAR LOCATION: Cell outer membrane
CC {ECO:0000269|PubMed:24257754}; Lipid-anchor {ECO:0000255|PROSITE-
CC ProRule:PRU00303, ECO:0000269|PubMed:24257754}.
CC -!- SIMILARITY: Belongs to the polysaccharide lyase 5 family.
CC {ECO:0000305}.
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DR EMBL; AM743169; CAQ45011.1; -; Genomic_DNA.
DR RefSeq; WP_012479599.1; NC_010943.1.
DR PDB; 7FHU; X-ray; 2.43 A; A/B=22-331.
DR PDB; 7FHV; X-ray; 2.28 A; A/B=22-331.
DR PDB; 7FHW; X-ray; 2.06 A; A/B=22-331.
DR PDB; 7FHX; X-ray; 2.63 A; A/B=22-331.
DR PDB; 7FHY; X-ray; 2.20 A; A/B=22-331.
DR PDB; 7FHZ; X-ray; 2.50 A; A=22-330.
DR PDB; 7FI0; X-ray; 2.31 A; A=22-331.
DR PDB; 7FI1; X-ray; 2.43 A; A=22-331.
DR PDB; 7FI2; X-ray; 2.60 A; A=22-331.
DR PDBsum; 7FHU; -.
DR PDBsum; 7FHV; -.
DR PDBsum; 7FHW; -.
DR PDBsum; 7FHX; -.
DR PDBsum; 7FHY; -.
DR PDBsum; 7FHZ; -.
DR PDBsum; 7FI0; -.
DR PDBsum; 7FI1; -.
DR PDBsum; 7FI2; -.
DR AlphaFoldDB; B2FHL8; -.
DR SMR; B2FHL8; -.
DR STRING; 522373.Smlt1473; -.
DR CAZy; PL5; Polysaccharide Lyase Family 5.
DR EnsemblBacteria; CAQ45011; CAQ45011; Smlt1473.
DR KEGG; sml:Smlt1473; -.
DR PATRIC; fig|522373.3.peg.1413; -.
DR eggNOG; ENOG502ZAMJ; Bacteria.
DR HOGENOM; CLU_064286_0_0_6; -.
DR OMA; AAWSVMA; -.
DR OrthoDB; 804604at2; -.
DR Proteomes; UP000008840; Chromosome.
DR GO; GO:0009279; C:cell outer membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0042597; C:periplasmic space; IEA:InterPro.
DR GO; GO:0033994; F:glucuronan lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0030340; F:hyaluronate lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0045135; F:poly(beta-D-mannuronate) lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.50.10.100; -; 1.
DR InterPro; IPR008397; Alginate_lyase_dom.
DR InterPro; IPR008929; Chondroitin_lyas.
DR Pfam; PF05426; Alginate_lyase; 1.
DR SUPFAM; SSF48230; SSF48230; 1.
DR PROSITE; PS51257; PROKAR_LIPOPROTEIN; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cell outer membrane; Lipoprotein;
KW Lyase; Membrane; Palmitate; Polysaccharide degradation; Reference proteome;
KW Signal.
FT SIGNAL 1..22
FT /evidence="ECO:0000305"
FT CHAIN 23..331
FT /note="Polysaccharide lyase"
FT /id="PRO_5000341778"
FT LIPID 23
FT /note="N-palmitoyl cysteine"
FT /evidence="ECO:0000305"
FT LIPID 23
FT /note="S-diacylglycerol cysteine"
FT /evidence="ECO:0000305"
FT MUTAGEN 23
FT /note="C->F: Impairs subcellular location since the protein
FT mutant is completely retained in the cytosol. Loss of
FT extracellular lyase activity in mutant cells."
FT /evidence="ECO:0000269|PubMed:24257754"
FT MUTAGEN 167
FT /note="N->L: Less than 0.5% of wild-type activity against
FT all substrates."
FT /evidence="ECO:0000269|PubMed:24257754"
FT MUTAGEN 168
FT /note="H->A: Retains 10% of activity toward poly-GlcA, but
FT shows less than 0.5% of wild-type activity against poly-
FT ManA and HA."
FT /evidence="ECO:0000269|PubMed:24257754"
FT MUTAGEN 171
FT /note="W->A: Displays nearly eliminated HA activity, while
FT increasing poly-ManA and poly-GlcA activity by at least
FT 35%."
FT /evidence="ECO:0000269|PubMed:24808176"
FT MUTAGEN 215
FT /note="R->L: Less than 0.5% of wild-type activity against
FT all substrates."
FT /evidence="ECO:0000269|PubMed:24257754"
FT MUTAGEN 221
FT /note="H->F: Increases activity and specificity toward
FT poly-ManA."
FT /evidence="ECO:0000269|PubMed:24808176"
FT MUTAGEN 222
FT /note="Y->F: Less than 0.5% of wild-type activity against
FT all substrates."
FT /evidence="ECO:0000269|PubMed:24257754"
FT MUTAGEN 312
FT /note="R->L: Increases activity and specificity toward
FT poly-GlcA."
FT /evidence="ECO:0000269|PubMed:24808176"
FT STRAND 31..33
FT /evidence="ECO:0007829|PDB:7FHZ"
FT HELIX 49..79
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 82..97
FT /evidence="ECO:0007829|PDB:7FHW"
FT TURN 98..101
FT /evidence="ECO:0007829|PDB:7FHW"
FT STRAND 103..105
FT /evidence="ECO:0007829|PDB:7FHZ"
FT STRAND 108..111
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 112..131
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 132..134
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 137..158
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 167..182
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 186..202
FT /evidence="ECO:0007829|PDB:7FHW"
FT TURN 210..214
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 216..218
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 219..239
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 244..246
FT /evidence="ECO:0007829|PDB:7FHU"
FT HELIX 247..264
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 267..273
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 286..292
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 295..297
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 300..303
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 311..313
FT /evidence="ECO:0007829|PDB:7FHW"
FT HELIX 317..323
FT /evidence="ECO:0007829|PDB:7FHW"
FT STRAND 325..328
FT /evidence="ECO:0007829|PDB:7FHW"
SQ SEQUENCE 331 AA; 36696 MW; B3D40DDFE2714589 CRC64;
MSLPLRLALL PTLLASASAF AACPAPPPGQ PDIRAIGYYT DKAGSVIDPA LQQQNKDATA
PLDRYAADVA RMSDDYLRNG DPAAAQCTLS WLGAWADDGA MLGQMIRVNN DQSFYMRQWM
LDAVAMAYLK VHDQANPQQR ARIDPWLQKL ARANLAYWDN PKRRRNNHYY WGGLGVLATG
LATDDDALWQ AGHAAFQKGI DDIQDDGSLP LEMARGQRAL HYHDYALAPL VMMAELARLR
GQDWYASRNH AIDRLARRVI EGSRDPAWFN QHTGAAQLPL QASGWVEFYR LRSPDGGVFD
AAHARGPFHS PRLGGDLTLM ATHGIVRTPL R
