CNL_CLINE
ID CNL_CLINE Reviewed; 149 AA.
AC B2ZRS9;
DT 16-MAR-2016, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-2008, sequence version 1.
DT 03-AUG-2022, entry version 46.
DE RecName: Full=Ricin B-like lectin {ECO:0000303|PubMed:19100814};
DE AltName: Full=Clitocybe nebularis lectin {ECO:0000303|PubMed:19100814};
DE Short=CNL {ECO:0000303|PubMed:19100814};
OS Clitocybe nebularis (Clouded agaric) (Lepista nebularis).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC Agaricomycetidae; Agaricales; Tricholomataceae; Clitocybe.
OX NCBI_TaxID=117024 {ECO:0000312|EMBL:ACD47153.1};
RN [1] {ECO:0000312|EMBL:ACD47153.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA], PROTEIN SEQUENCE OF 3-32; 41-70;
RP 74-105; 109-114; 117-136 AND 142-149, FUNCTION, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, SUBUNIT, PTM, MASS SPECTROMETRY, AND
RP ACETYLATION AT SER-2.
RC STRAIN=Vrh2004 {ECO:0000312|EMBL:ACD47153.1};
RC TISSUE=Fruiting body {ECO:0000312|EMBL:ACD47153.1};
RX PubMed=19100814; DOI=10.1016/j.bbagen.2008.11.006;
RA Pohleven J., Obermajer N., Sabotic J., Anzlovar S., Sepcic K., Kos J.,
RA Kralj B., Strukelj B., Brzin J.;
RT "Purification, characterization and cloning of a ricin B-like lectin from
RT mushroom Clitocybe nebularis with antiproliferative activity against human
RT leukemic T cells.";
RL Biochim. Biophys. Acta 1790:173-181(2009).
RN [2] {ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD, ECO:0007744|PDB:3NBE}
RP PARTIAL PROTEIN SEQUENCE, X-RAY CRYSTALLOGRAPHY (1.01 ANGSTROMS) OF 2-149
RP IN COMPLEX WITH LACTOSE AND LDN, FUNCTION, SUBSTRATE SPECIFICITY, SUBUNIT,
RP DOMAIN, MASS SPECTROMETRY, CIRCULAR DICHROISM ANALYSIS, AND MUTAGENESIS OF
RP ASP-21; LEU-55 AND ASN-111.
RC TISSUE=Fruiting body {ECO:0000269|PubMed:22298779};
RX PubMed=22298779; DOI=10.1074/jbc.m111.317263;
RA Pohleven J., Renko M., Magister S., Smith D.F., Kunzler M., Strukelj B.,
RA Turk D., Kos J., Sabotic J.;
RT "Bivalent carbohydrate binding is required for biological activity of
RT Clitocybe nebularis lectin (CNL), the N,N'-diacetyllactosediamine
RT (GalNAcbeta1-4GlcNAc, LacdiNAc)-specific lectin from basidiomycete C.
RT nebularis.";
RL J. Biol. Chem. 287:10602-10612(2012).
RN [3]
RP FUNCTION, MASS SPECTROMETRY, AND BIOTECHNOLOGY.
RX PubMed=21556921; DOI=10.1007/s00253-011-3236-0;
RA Pohleven J., Brzin J., Vrabec L., Leonardi A., Cokl A., Strukelj B.,
RA Kos J., Sabotic J.;
RT "Basidiomycete Clitocybe nebularis is rich in lectins with insecticidal
RT activities.";
RL Appl. Microbiol. Biotechnol. 91:1141-1148(2011).
RN [4]
RP FUNCTION.
RX PubMed=22044067; DOI=10.1111/j.1365-2567.2011.03500.x;
RA Svajger U., Pohleven J., Kos J., Strukelj B., Jeras M.;
RT "CNL, a ricin B-like lectin from mushroom Clitocybe nebularis, induces
RT maturation and activation of dendritic cells via the toll-like receptor 4
RT pathway.";
RL Immunology 134:409-418(2011).
RN [5]
RP FUNCTION.
RX PubMed=21486374; DOI=10.1111/j.1365-294x.2011.05093.x;
RA Bleuler-Martinez S., Butschi A., Garbani M., Waelti M.A., Wohlschlager T.,
RA Potthoff E., Sabotic J., Pohleven J., Luethy P., Hengartner M.O., Aebi M.,
RA Kuenzler M.;
RT "A lectin-mediated resistance of higher fungi against predators and
RT parasites.";
RL Mol. Ecol. 20:3056-3070(2011).
CC -!- FUNCTION: Lectin specific for terminal, non-reducing N-
CC acetylgalactosamine (Gal-NAc)-containing carbohydrates including N,N'-
CC diacetyllactosediamine/LDN (GalNAcbeta1-4GlcNAc, LacdiNAc). Specific
CC also for carbohydrates containing N-acetylglucosamine (-GlcNAc) or N-
CC acetyllactosamine (-Galbeta1-4GlcNAc) at the reducing end. Agglutinates
CC human blood group A, AB, B and O erythrocytes with a strong preference
CC for group A. Agglutinates bovine erythrocytes with a very low
CC specificity (PubMed:19100814, PubMed:22298779). Binds carbohydrates
CC bivalently, which is required for its biological activity
CC (PubMed:22298779). Exhibits insecticidal activity against the fruit fly
CC D.melanogaster, mosquito A.aegypti, and amoebozoa A.castellanii. Has
CC anti-nutritional activity against Colorado potato beetle
CC L.decemlineata, and against worm C.elegans (PubMed:21556921,
CC PubMed:21486374). Has antiproliferative activity against human leukemic
CC T-cells (PubMed:19100814). Has an immunostimulatory effect on human
CC antigen-presenting dendritic cells, which are subsequently able to
CC induce efficient T-cell immune responses (PubMed:22044067).
CC {ECO:0000269|PubMed:19100814, ECO:0000269|PubMed:21486374,
CC ECO:0000269|PubMed:21556921, ECO:0000269|PubMed:22044067,
CC ECO:0000269|PubMed:22298779}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 6-6.5. Retains more than 70% of the maximum
CC agglutinating activity over the pH range 5-9.
CC {ECO:0000269|PubMed:19100814};
CC Temperature dependence:
CC Unchanged hemagglutinating activity after 30 minutes incubation at
CC temperatures up to 50 degrees Celsius, then at higher temperatures
CC activity decreases. 14% of the initial activity is retained even
CC after heating at 100 degrees Celsius. {ECO:0000269|PubMed:19100814};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:19100814,
CC ECO:0000269|PubMed:22298779}.
CC -!- DOMAIN: Contains a single ricin B lectin domain with a characteristic
CC beta-trefoil fold consisting of three repeated subdomains each
CC containing a more or less conserved QXW motif.
CC {ECO:0000305|PubMed:22298779}.
CC -!- PTM: The N-terminus is blocked. {ECO:0000269|PubMed:19100814}.
CC -!- MASS SPECTROMETRY: Mass=15903.5; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:19100814};
CC -!- MASS SPECTROMETRY: Mass=15992; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:22298779};
CC -!- MASS SPECTROMETRY: Mass=15903; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:21556921};
CC -!- BIOTECHNOLOGY: Potential for use as natural insecticide.
CC {ECO:0000303|PubMed:21556921}.
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DR EMBL; EU682006; ACD47153.1; -; Genomic_DNA.
DR EMBL; FJ477895; ACK56062.1; -; mRNA.
DR PDB; 3NBC; X-ray; 1.01 A; A/B=2-149.
DR PDB; 3NBD; X-ray; 1.15 A; A/B=2-149.
DR PDB; 3NBE; X-ray; 1.86 A; A/B=2-149.
DR PDBsum; 3NBC; -.
DR PDBsum; 3NBD; -.
DR PDBsum; 3NBE; -.
DR AlphaFoldDB; B2ZRS9; -.
DR SMR; B2ZRS9; -.
DR UniLectin; B2ZRS9; -.
DR iPTMnet; B2ZRS9; -.
DR EvolutionaryTrace; B2ZRS9; -.
DR GO; GO:0030246; F:carbohydrate binding; IEA:UniProtKB-KW.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR InterPro; IPR035992; Ricin_B-like_lectins.
DR InterPro; IPR000772; Ricin_B_lectin.
DR Pfam; PF14200; RicinB_lectin_2; 1.
DR SUPFAM; SSF50370; SSF50370; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Direct protein sequencing; Lectin.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:19100814"
FT CHAIN 2..149
FT /note="Ricin B-like lectin"
FT /id="PRO_0000435793"
FT REGION 110..112
FT /note="Involved in dimerization"
FT /evidence="ECO:0000269|PubMed:22298779,
FT ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD"
FT BINDING 21
FT /ligand="a carbohydrate"
FT /ligand_id="ChEBI:CHEBI:16646"
FT /evidence="ECO:0000269|PubMed:22298779,
FT ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD"
FT BINDING 24
FT /ligand="a carbohydrate"
FT /ligand_id="ChEBI:CHEBI:16646"
FT /evidence="ECO:0000269|PubMed:22298779,
FT ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD"
FT BINDING 39
FT /ligand="a carbohydrate"
FT /ligand_id="ChEBI:CHEBI:16646"
FT /evidence="ECO:0000269|PubMed:22298779,
FT ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD"
FT BINDING 47
FT /ligand="a carbohydrate"
FT /ligand_id="ChEBI:CHEBI:16646"
FT /evidence="ECO:0000269|PubMed:22298779,
FT ECO:0007744|PDB:3NBC, ECO:0007744|PDB:3NBD"
FT MOD_RES 2
FT /note="N-acetylserine"
FT /evidence="ECO:0000269|PubMed:19100814"
FT MUTAGEN 21
FT /note="D->R: No sugar-binding, hemagglutination of human
FT group A erythrocytes nor toxicity to Jurkat leukemia
FT cells."
FT /evidence="ECO:0000269|PubMed:22298779"
FT MUTAGEN 55
FT /note="L->R: No effect on sugar-binding, but loss of
FT dimerization, hemagglutination of human group A
FT erythrocytes and toxicity to Jurkat leukemia cells; when
FT associated with D-111."
FT /evidence="ECO:0000269|PubMed:22298779"
FT MUTAGEN 55
FT /note="L->W: No effect on sugar-binding, but loss of
FT dimerization, hemagglutination of human group A
FT erythrocytes and toxicity to Jurkat leukemia cells; when
FT associated with D-111."
FT /evidence="ECO:0000269|PubMed:22298779"
FT MUTAGEN 111
FT /note="N->D: No effect on sugar-binding, hemagglutination
FT of human group A erythrocytes and dimerization, but reduced
FT toxicity to Jurkat leukemia cells. No effect on sugar-
FT binding, but loss of dimerization, hemagglutination of
FT human group A erythrocytes and toxicity to Jurkat leukemia
FT cells; when associated with R-55. No effect on sugar-
FT binding, but loss of dimerization, hemagglutination of
FT human group A erythrocytes and toxicity to Jurkat leukemia
FT cells; when associated with W-55."
FT /evidence="ECO:0000269|PubMed:22298779"
FT CONFLICT 31
FT /note="T -> V (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 32
FT /note="L -> I (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 5..14
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 19..22
FT /evidence="ECO:0007829|PDB:3NBC"
FT HELIX 23..25
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 31..36
FT /evidence="ECO:0007829|PDB:3NBC"
FT TURN 41..43
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 50..54
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 58..68
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 71..74
FT /evidence="ECO:0007829|PDB:3NBC"
FT HELIX 75..77
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 84..90
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 94..98
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 105..108
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 112..117
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 128..131
FT /evidence="ECO:0007829|PDB:3NBC"
FT HELIX 137..139
FT /evidence="ECO:0007829|PDB:3NBC"
FT STRAND 141..145
FT /evidence="ECO:0007829|PDB:3NBC"
SQ SEQUENCE 149 AA; 15993 MW; 28E71535B79818CB CRC64;
MSITPGTYNI TNVAYTNRLI DLTGSNPAEN TLIIGHHLNK TPSGYGNQQW TLVQLPHTTI
YTMQAVNPQS YVRVRDDNLV DGAALVGSQQ PTPVSIESAG NSGQFRIKIP NLGLALTLPS
DANSTPIVLG EVDETSTNQL WAFESVSAV
