TCSL1_PAESO
ID TCSL1_PAESO Reviewed; 2364 AA.
AC Q46342;
DT 07-OCT-2020, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 97.
DE RecName: Full=Cytotoxin-L {ECO:0000303|PubMed:7642137};
DE EC=3.4.22.- {ECO:0000269|PubMed:17334356};
DE AltName: Full=Lethal toxin {ECO:0000303|PubMed:8626575, ECO:0000303|PubMed:8626586};
DE Short=LT {ECO:0000303|PubMed:8626575, ECO:0000303|PubMed:8626586};
DE Contains:
DE RecName: Full=Glucosyltransferase TcsL {ECO:0000305};
DE EC=2.4.1.- {ECO:0000269|PubMed:17901056, ECO:0000269|PubMed:19744486, ECO:0000269|PubMed:24905543, ECO:0000269|PubMed:8626575, ECO:0000269|PubMed:8626586, ECO:0000269|PubMed:8858106, ECO:0000269|PubMed:9632667};
GN Name=tcsL {ECO:0000303|PubMed:17910886, ECO:0000303|PubMed:27303685};
OS Paeniclostridium sordellii (Clostridium sordellii).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Peptostreptococcaceae;
OC Paeniclostridium.
OX NCBI_TaxID=1505;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=6018 / IP82;
RX PubMed=7642137; DOI=10.1016/0378-1119(95)00263-6;
RA Green G.A., Schue V., Monteil H.;
RT "Cloning and characterization of the cytotoxin L-encoding gene of
RT Clostridium sordellii: homology with Clostridium difficile cytotoxin B.";
RL Gene 161:57-61(1995).
RN [2]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=8858106; DOI=10.1006/bbrc.1996.1470;
RA Hofmann F., Rex G., Aktories K., Just I.;
RT "The ras-related protein Ral is monoglucosylated by Clostridium sordellii
RT lethal toxin.";
RL Biochem. Biophys. Res. Commun. 227:77-81(1996).
RN [3]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL), AND COFACTOR (GLUCOSYLTRANSFERASE TCSL).
RX PubMed=8626575; DOI=10.1074/jbc.271.17.10149;
RA Just I., Selzer J., Hofmann F., Green G.A., Aktories K.;
RT "Inactivation of Ras by Clostridium sordellii lethal toxin-catalyzed
RT glucosylation.";
RL J. Biol. Chem. 271:10149-10153(1996).
RN [4]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL), AND SUBCELLULAR LOCATION (GLUCOSYLTRANSFERASE
RP TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=8626586; DOI=10.1074/jbc.271.17.10217;
RA Popoff M.R., Chaves-Olarte E., Lemichez E., von Eichel-Streiber C.,
RA Thelestam M., Chardin P., Cussac D., Antonny B., Chavrier P., Flatau G.,
RA Giry M., de Gunzburg J., Boquet P.;
RT "Ras, Rap, and Rac small GTP-binding proteins are targets for Clostridium
RT sordellii lethal toxin glucosylation.";
RL J. Biol. Chem. 271:10217-10224(1996).
RN [5]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=9632667; DOI=10.1074/jbc.273.26.16134;
RA Herrmann C., Ahmadian M.R., Hofmann F., Just I.;
RT "Functional consequences of monoglucosylation of Ha-Ras at effector domain
RT amino acid threonine 35.";
RL J. Biol. Chem. 273:16134-16139(1998).
RN [6]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL).
RX PubMed=17910886; DOI=10.1016/j.bbrc.2007.09.073;
RA Voth D.E., Ballard J.D.;
RT "Critical intermediate steps in Clostridium sordellii lethal toxin-induced
RT apoptosis.";
RL Biochem. Biophys. Res. Commun. 363:959-964(2007).
RN [7]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=17901056; DOI=10.1074/jbc.m703138200;
RA Jank T., Giesemann T., Aktories K.;
RT "Clostridium difficile glucosyltransferase toxin B-essential amino acids
RT for substrate binding.";
RL J. Biol. Chem. 282:35222-35231(2007).
RN [8]
RP FUNCTION (CYTOTOXIN-L), ACTIVITY REGULATION (CYTOTOXIN-L), AND PROTEOLYTIC
RP CLEAVAGE (CYTOTOXIN-L).
RX PubMed=17334356; DOI=10.1038/nature05622;
RA Reineke J., Tenzer S., Rupnik M., Koschinski A., Hasselmayer O.,
RA Schrattenholz A., Schild H., von Eichel-Streiber C.;
RT "Autocatalytic cleavage of Clostridium difficile toxin B.";
RL Nature 446:415-419(2007).
RN [9]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=19744486; DOI=10.1016/j.febslet.2009.09.006;
RA Huelsenbeck S.C., Klose I., Reichenbach M., Huelsenbeck J., Genth H.;
RT "Distinct kinetics of (H/K/N)Ras glucosylation and Rac1 glucosylation
RT catalysed by Clostridium sordellii lethal toxin.";
RL FEBS Lett. 583:3133-3139(2009).
RN [10]
RP FUNCTION.
RX PubMed=19527792; DOI=10.1016/j.anaerobe.2009.06.002;
RA Hao Y., Senn T., Opp J.S., Young V.B., Thiele T., Srinivas G., Huang S.K.,
RA Aronoff D.M.;
RT "Lethal toxin is a critical determinant of rapid mortality in rodent models
RT of Clostridium sordellii endometritis.";
RL Anaerobe 16:155-160(2010).
RN [11]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=24905543; DOI=10.1111/cmi.12321;
RA Genth H., Pauillac S., Schelle I., Bouvet P., Bouchier C.,
RA Varela-Chavez C., Just I., Popoff M.R.;
RT "Haemorrhagic toxin and lethal toxin from Clostridium sordellii strain
RT vpi9048: molecular characterization and comparative analysis of substrate
RT specificity of the large clostridial glucosylating toxins.";
RL Cell. Microbiol. 16:1706-1721(2014).
RN [12]
RP FUNCTION, CATALYTIC ACTIVITY (GLUCOSYLTRANSFERASE TCSL), AND MUTAGENESIS OF
RP 286-ASP--ASP-288.
RX PubMed=24919149; DOI=10.1038/nature13449;
RA Xu H., Yang J., Gao W., Li L., Li P., Zhang L., Gong Y.N., Peng X.,
RA Xi J.J., Chen S., Wang F., Shao F.;
RT "Innate immune sensing of bacterial modifications of Rho GTPases by the
RT Pyrin inflammasome.";
RL Nature 513:237-241(2014).
RN [13]
RP SUBCELLULAR LOCATION (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=25882477; DOI=10.1111/cmi.12449;
RA Varela Chavez C., Hoos S., Haustant G.M., Chenal A., England P.,
RA Blondel A., Pauillac S., Lacy D.B., Popoff M.R.;
RT "The catalytic domains of Clostridium sordellii lethal toxin and related
RT large clostridial glucosylating toxins specifically recognize the
RT negatively charged phospholipids phosphatidylserine and phosphatidic
RT acid.";
RL Cell. Microbiol. 17:1477-1493(2015).
RN [14]
RP FUNCTION (CYTOTOXIN-L), FUNCTION (GLUCOSYLTRANSFERASE TCSL), CATALYTIC
RP ACTIVITY (GLUCOSYLTRANSFERASE TCSL), PROTEOLYTIC CLEAVAGE (CYTOTOXIN-L),
RP SUBCELLULAR LOCATION (GLUCOSYLTRANSFERASE TCSL), AND MUTAGENESIS OF
RP 17-PHE-ARG-18; 286-ASP--ASP-288 AND CYS-698.
RC STRAIN=JGS6382;
RX PubMed=27303685; DOI=10.1128/msphere.00012-15;
RA Craven R., Lacy D.B.;
RT "Clostridium sordellii lethal-toxin autoprocessing and membrane
RT localization activities drive GTPase glucosylation profiles in endothelial
RT cells.";
RL MSphere 1:0-0(2016).
RN [15]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL), SUBCELLULAR LOCATION (GLUCOSYLTRANSFERASE
RP TCSL), DOMAIN (GLUCOSYLTRANSFERASE TCSL), AND MUTAGENESIS OF GLN-10;
RP LYS-11; TYR-14; VAL-15; LYS-16; PHE-17; ARG-18; GLN-20; SER-38; ARG-68;
RP TYR-78 AND 286-ASP--ASP-288.
RX PubMed=27023605; DOI=10.3390/toxins8040090;
RA Varela Chavez C., Haustant G.M., Baron B., England P., Chenal A.,
RA Pauillac S., Blondel A., Popoff M.R.;
RT "The tip of the four N-terminal alpha-helices of Clostridium sordellii
RT lethal toxin contains the interaction site with membrane phosphatidylserine
RT facilitating small GTPases glucosylation.";
RL Toxins 8:90-90(2016).
RN [16]
RP COFACTOR (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=27089365; DOI=10.3390/toxins8040109;
RA Genth H., Schelle I., Just I.;
RT "Metal ion activation of clostridium sordellii lethal toxin and Clostridium
RT difficile toxin B.";
RL Toxins 8:109-109(2016).
RN [17]
RP FUNCTION (GLUCOSYLTRANSFERASE TCSL), AND CATALYTIC ACTIVITY
RP (GLUCOSYLTRANSFERASE TCSL).
RC STRAIN=6018 / IP82;
RX PubMed=30622517; DOI=10.3389/fmicb.2018.03078;
RA Genth H., Junemann J., Laemmerhirt C.M., Luecke A.C., Schelle I., Just I.,
RA Gerhard R., Pich A.;
RT "Difference in mono-O-glucosylation of Ras subtype GTPases between toxin A
RT and toxin B from Clostridioides difficile strain 10463 and lethal toxin
RT from Clostridium sordellii strain 6018.";
RL Front. Microbiol. 9:3078-3078(2018).
RN [18]
RP REVIEW.
RX PubMed=29146177; DOI=10.1016/j.toxicon.2017.11.003;
RA Popoff M.R.;
RT "Clostridium difficile and Clostridium sordellii toxins, proinflammatory
RT versus anti-inflammatory response.";
RL Toxicon 149:54-64(2018).
RN [19]
RP INTERACTION WITH HOST SEMA6A AND SEMA6B (CYTOTOXIN-L).
RC STRAIN=6018 / IP82;
RX PubMed=32302524; DOI=10.1016/j.chom.2020.03.007;
RA Tian S., Liu Y., Wu H., Liu H., Zeng J., Choi M.Y., Chen H., Gerhard R.,
RA Dong M.;
RT "Genome-wide CRISPR screen identifies semaphorin 6A and 6B as receptors for
RT Paeniclostridium sordellii toxin TcsL.";
RL Cell Host Microbe 27:782-792(2020).
RN [20] {ECO:0007744|PDB:2VL8}
RP X-RAY CRYSTALLOGRAPHY (2.31 ANGSTROMS) OF 1-546 IN COMPLEX WITH CALCIUM AND
RP UDP.
RC STRAIN=6018 / IP82;
RX PubMed=18505687; DOI=10.1016/j.febslet.2008.05.025;
RA Jank T., Ziegler M.O., Schulz G.E., Aktories K.;
RT "Inhibition of the glucosyltransferase activity of clostridial Rho/Ras-
RT glucosylating toxins by castanospermine.";
RL FEBS Lett. 582:2277-2282(2008).
RN [21] {ECO:0007744|PDB:2VKD, ECO:0007744|PDB:2VKH}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 1-546 IN COMPLEX WITH
RP UDP-ALPHA-D-GLUCOSE AND MANGANESE.
RC STRAIN=6018 / IP82;
RX PubMed=18325534; DOI=10.1016/j.jmb.2007.12.065;
RA Ziegler M.O., Jank T., Aktories K., Schulz G.E.;
RT "Conformational changes and reaction of clostridial glycosylating toxins.";
RL J. Mol. Biol. 377:1346-1356(2008).
CC -!- FUNCTION: [Cytotoxin-L]: Precursor of a cytotoxin that targets the
CC vascular endothelium, inducing an anti-inflammatory effect and
CC resulting in lethal toxic shock syndrome (PubMed:19527792,
CC PubMed:24919149, PubMed:29146177). TcsL constitutes the main toxin that
CC mediates the pathology of P.sordellii infection, an anaerobic Gram-
CC positive bacterium found in soil and in the gastrointestinal and
CC vaginal tracts of animals and humans; although the majority of carriers
CC are asymptomatic, pathogenic P.sordellii infections arise rapidly and
CC are highly lethal (PubMed:29146177). This form constitutes the
CC precursor of the toxin: it enters into host cells and mediates
CC autoprocessing to release the active toxin (Glucosyltransferase TcsL)
CC into the host cytosol (PubMed:32302524, PubMed:17334356,
CC PubMed:27303685). Targets vascular endothelium by binding to the
CC semaphorin proteins SEMA6A and SEMA6B, and enters host cells via
CC clathrin-mediated endocytosis (PubMed:32302524). Once entered into host
CC cells, acidification in the endosome promotes the membrane insertion of
CC the translocation region and formation of a pore, leading to
CC translocation of the GT44 and peptidase C80 domains across the
CC endosomal membrane (By similarity). This activates the peptidase C80
CC domain and autocatalytic processing, releasing the N-terminal part
CC (Glucosyltransferase TcsL), which constitutes the active part of the
CC toxin, in the cytosol (PubMed:17334356, PubMed:27303685).
CC {ECO:0000250|UniProtKB:P18177, ECO:0000269|PubMed:17334356,
CC ECO:0000269|PubMed:19527792, ECO:0000269|PubMed:24919149,
CC ECO:0000269|PubMed:27303685, ECO:0000269|PubMed:32302524,
CC ECO:0000303|PubMed:29146177}.
CC -!- FUNCTION: [Glucosyltransferase TcsL]: Active form of the toxin, which
CC is released into the host cytosol following autoprocessing and
CC inactivates small GTPases (PubMed:8626575, PubMed:8626586,
CC PubMed:9632667, PubMed:17901056, PubMed:19744486, PubMed:24905543,
CC PubMed:24919149, PubMed:27303685, PubMed:27023605, PubMed:30622517).
CC Acts by mediating monoglucosylation of small GTPases of the Ras (H-
CC Ras/HRAS, K-Ras/KRAS, N-Ras/NRAS and Ral/RALA) family in host cells at
CC the conserved threonine residue located in the switch I region ('Thr-
CC 37/35'), using UDP-alpha-D-glucose as the sugar donor (PubMed:8858106,
CC PubMed:8626575, PubMed:8626586, PubMed:9632667, PubMed:17901056,
CC PubMed:19744486, PubMed:24905543, PubMed:24919149, PubMed:27023605,
CC PubMed:30622517). Also able to catalyze monoglucosylation of some
CC members of the Rho family (Rac1 and Rap2A), but with less efficiency
CC than with Ras proteins (PubMed:8626586, PubMed:9632667,
CC PubMed:19744486, PubMed:24905543). Monoglucosylation of host small
CC GTPases completely prevents the recognition of the downstream effector,
CC blocking the GTPases in their inactive form and leading to apoptosis
CC (PubMed:8626586, PubMed:9632667, PubMed:17910886). Induces an anti-
CC inflammatory effect, mainly by inactivating Ras proteins which results
CC in blockage of the cell cycle and killing of immune cells
CC (PubMed:17910886, PubMed:24919149). The absence or moderate local
CC inflammatory response allows C.sordellii spreading in deep tissues,
CC production of toxin which is released in the general circulation and
CC causes a toxic shock syndrome (PubMed:24919149, PubMed:29146177).
CC {ECO:0000269|PubMed:17901056, ECO:0000269|PubMed:17910886,
CC ECO:0000269|PubMed:19744486, ECO:0000269|PubMed:24905543,
CC ECO:0000269|PubMed:24919149, ECO:0000269|PubMed:27023605,
CC ECO:0000269|PubMed:27303685, ECO:0000269|PubMed:30622517,
CC ECO:0000269|PubMed:8626575, ECO:0000269|PubMed:8626586,
CC ECO:0000269|PubMed:8858106, ECO:0000269|PubMed:9632667,
CC ECO:0000303|PubMed:29146177}.
CC -!- CATALYTIC ACTIVITY: [Glucosyltransferase TcsL]:
CC Reaction=L-threonyl-[protein] + UDP-alpha-D-glucose = 3-O-(alpha-D-
CC glucosyl)-L-threonyl-[protein] + H(+) + UDP; Xref=Rhea:RHEA:64684,
CC Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:16656, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30013, ChEBI:CHEBI:58223, ChEBI:CHEBI:58885,
CC ChEBI:CHEBI:156085; Evidence={ECO:0000269|PubMed:17901056,
CC ECO:0000269|PubMed:19744486, ECO:0000269|PubMed:24905543,
CC ECO:0000269|PubMed:24919149, ECO:0000269|PubMed:27023605,
CC ECO:0000269|PubMed:27303685, ECO:0000269|PubMed:30622517,
CC ECO:0000269|PubMed:8626575, ECO:0000269|PubMed:8626586,
CC ECO:0000269|PubMed:8858106, ECO:0000269|PubMed:9632667};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:64685;
CC Evidence={ECO:0000269|PubMed:17901056, ECO:0000269|PubMed:19744486,
CC ECO:0000269|PubMed:24905543, ECO:0000269|PubMed:24919149,
CC ECO:0000269|PubMed:27023605, ECO:0000269|PubMed:27303685,
CC ECO:0000269|PubMed:30622517, ECO:0000269|PubMed:8626575,
CC ECO:0000269|PubMed:8626586, ECO:0000269|PubMed:8858106,
CC ECO:0000269|PubMed:9632667};
CC -!- COFACTOR: [Cytotoxin-L]:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000250|UniProtKB:P18177};
CC Note=Binds 1 Zn(2+) ion per subunit. Zn(2+) is required for
CC autocatalytic cleavage. {ECO:0000250|UniProtKB:P18177};
CC -!- COFACTOR: [Glucosyltransferase TcsL]:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:27089365, ECO:0000269|PubMed:8626575};
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:27089365};
CC Note=Has higher activity with Mn(2+), but most likely uses Mg(2+) in
CC host cells (PubMed:27089365). Mn(2+) or Mg(2+) are required for
CC glucosyltransferase activity (PubMed:8626575, PubMed:27089365).
CC {ECO:0000269|PubMed:27089365, ECO:0000269|PubMed:8626575};
CC -!- ACTIVITY REGULATION: [Cytotoxin-L]: Protease activity is activated upon
CC binding to 1D-myo-inositol hexakisphosphate (InsP6), which induces
CC conformational reorganization. {ECO:0000269|PubMed:17334356}.
CC -!- SUBUNIT: [Cytotoxin-L]: Homomultimer; forms an inactive homomultimer at
CC pH 8, which dissociates at pH 4, leading to cytotoxicity (By
CC similarity). Interacts with host SEMA6A; interaction promotes toxin
CC entry into host cell (PubMed:32302524). Interacts with host SEMA6B;
CC interaction promotes toxin entry into host cell (PubMed:32302524).
CC {ECO:0000250|UniProtKB:T0D3N5, ECO:0000269|PubMed:32302524}.
CC -!- SUBCELLULAR LOCATION: [Cytotoxin-L]: Secreted
CC {ECO:0000250|UniProtKB:P18177}. Host endosome membrane
CC {ECO:0000250|UniProtKB:P18177}. Note=Secreted from P.sordellii cell
CC into the extracellular environment via help of holin-like protein
CC TcdE/UtxA (By similarity). Binds to the cell surface receptors via the
CC receptor-binding region and enters the cells via clathrin-mediated
CC endocytosis (By similarity). Acidification in the endosome triggers
CC conformational changes that promote the membrane insertion of the
CC translocation region, allowing formation of a pore, leading to
CC translocation of the GT44 and peptidase C80 domains across the
CC endosomal membrane (By similarity). 1D-myo-inositol hexakisphosphate-
CC binding (InsP6) activates the peptidase C80 domain and autoprocessing,
CC generating the Glucosyltransferase TcsL form, which is released in the
CC host cytosol (PubMed:17334356). {ECO:0000250|UniProtKB:P18177,
CC ECO:0000269|PubMed:17334356}.
CC -!- SUBCELLULAR LOCATION: [Glucosyltransferase TcsL]: Host cytoplasm, host
CC cytosol {ECO:0000305|PubMed:8626586}. Host cell membrane
CC {ECO:0000269|PubMed:27023605, ECO:0000269|PubMed:27303685}; Peripheral
CC membrane protein {ECO:0000269|PubMed:27023605}; Cytoplasmic side
CC {ECO:0000269|PubMed:27023605}. Note=Binding to phospholipids, such as
CC phosphatidylserine and phosphatidic acid promotes localization to the
CC inner face of the cell membrane close to its membrane anchored
CC substrates (small GTPases). {ECO:0000269|PubMed:25882477,
CC ECO:0000269|PubMed:27023605}.
CC -!- DOMAIN: [Glucosyltransferase TcsL]: Consists of 4 functional domains:
CC (1) the N-terminal GT44 domain (glucosyltransferase, also named GTD),
CC which mediates glucosylation of host small GTPases, (2) an
CC autoprocessing region that catalyzes autoprocessing to release the N-
CC terminal GT44 domain in the host cytosol, (3) the translocation region
CC that forms a pore to promote translocation of the GT44 and peptidase
CC C80 domains across the endosomal membrane and (4) the receptor-binding
CC (CROPS) region that mediates binding to host cells and contribute to
CC entry into cells. {ECO:0000303|PubMed:29146177}.
CC -!- DOMAIN: [Cytotoxin-L]: The receptor-binding (CROPS) region is dynamic
CC and can have open and closed conformations depending of the pH: has an
CC open conformation at endosomal pH and a closed conformation at neutral
CC pH. {ECO:0000250|UniProtKB:P18177}.
CC -!- DOMAIN: [Cytotoxin-L]: The cell wall-binding repeats bind
CC carbohydrates, probably contributing to entry into cells.
CC {ECO:0000250|UniProtKB:P16154}.
CC -!- DOMAIN: [Glucosyltransferase TcsL]: The four-helical bundle region
CC mediates binding to phospholipids, such as phosphatidylserine and
CC phosphatidic acid (PubMed:27023605). This promotes localization to the
CC inner face of the cell membrane close to small GTPases
CC (PubMed:27023605). {ECO:0000269|PubMed:27023605}.
CC -!- PTM: [Cytotoxin-L]: Undergoes autocatalytic cleavage to release the N-
CC terminal part (Glucosyltransferase TcsL), which constitutes the active
CC part of the toxin, in the host cytosol (PubMed:17334356,
CC PubMed:27303685). 1D-myo-inositol hexakisphosphate-binding (InsP6)
CC activates the peptidase C80 domain and promotes autoprocessing
CC (PubMed:17334356). {ECO:0000269|PubMed:17334356,
CC ECO:0000269|PubMed:27303685}.
CC -!- SIMILARITY: Belongs to the clostridial glucosylating toxin (LCGT)
CC family. {ECO:0000305}.
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DR EMBL; X82638; CAA57959.1; -; Genomic_DNA.
DR PIR; I40884; I40884.
DR PDB; 2VKD; X-ray; 2.53 A; A/B/C=1-546.
DR PDB; 2VKH; X-ray; 2.30 A; A/B/C=1-546.
DR PDB; 2VL8; X-ray; 2.31 A; A/B/C=1-546.
DR PDBsum; 2VKD; -.
DR PDBsum; 2VKH; -.
DR PDBsum; 2VL8; -.
DR AlphaFoldDB; Q46342; -.
DR SMR; Q46342; -.
DR IntAct; Q46342; 4.
DR MINT; Q46342; -.
DR CAZy; GT44; Glycosyltransferase Family 44.
DR TCDB; 1.C.57.1.3; the clostridial cytotoxin (cct) family.
DR BRENDA; 2.4.1.B62; 1514.
DR EvolutionaryTrace; Q46342; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0044164; C:host cell cytosol; IEA:UniProtKB-SubCell.
DR GO; GO:0044175; C:host cell endosome membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0020002; C:host cell plasma membrane; IDA:UniProtKB.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-KW.
DR GO; GO:0008234; F:cysteine-type peptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0016757; F:glycosyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0046789; F:host cell surface receptor binding; IPI:UniProtKB.
DR GO; GO:0008289; F:lipid binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008233; F:peptidase activity; IDA:UniProtKB.
DR GO; GO:0005543; F:phospholipid binding; IDA:UniProtKB.
DR GO; GO:0090729; F:toxin activity; IDA:UniProtKB.
DR GO; GO:0035251; F:UDP-glucosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0034260; P:negative regulation of GTPase activity; IDA:UniProtKB.
DR GO; GO:0046580; P:negative regulation of Ras protein signal transduction; IDA:UniProtKB.
DR GO; GO:0016540; P:protein autoprocessing; IDA:UniProtKB.
DR GO; GO:0018243; P:protein O-linked glycosylation via threonine; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.11050; -; 1.
DR InterPro; IPR018337; Cell_wall/Cho-bd_repeat.
DR InterPro; IPR020974; CPD_dom.
DR InterPro; IPR038383; CPD_dom_sf.
DR InterPro; IPR020972; Dermonecrotic/RTX_toxin_MLD.
DR InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR InterPro; IPR024770; TcdA/TcdB_cat.
DR InterPro; IPR024772; TcdA/TcdB_N.
DR InterPro; IPR024769; TcdA/TcdB_pore_forming.
DR Pfam; PF01473; Choline_bind_1; 5.
DR Pfam; PF11647; MLD; 1.
DR Pfam; PF11713; Peptidase_C80; 1.
DR Pfam; PF12919; TcdA_TcdB; 1.
DR Pfam; PF12920; TcdA_TcdB_pore; 1.
DR Pfam; PF12918; TcdB_N; 1.
DR SUPFAM; SSF53448; SSF53448; 1.
DR PROSITE; PS51771; CGT_MARTX_CPD; 1.
DR PROSITE; PS51170; CW; 20.
PE 1: Evidence at protein level;
KW 3D-structure; Autocatalytic cleavage; Glycosyltransferase;
KW Host cell membrane; Host cytoplasm; Host endosome; Host membrane;
KW Hydrolase; Lipid-binding; Magnesium; Manganese; Membrane; Metal-binding;
KW Protease; Repeat; Secreted; Thiol protease; Toxin; Transferase; Virulence;
KW Zinc.
FT CHAIN 1..2364
FT /note="Cytotoxin-L"
FT /id="PRO_0000451197"
FT CHAIN 1..543
FT /note="Glucosyltransferase TcsL"
FT /evidence="ECO:0000250|UniProtKB:P18177"
FT /id="PRO_0000451198"
FT DOMAIN 96..468
FT /note="GT44"
FT /evidence="ECO:0000255"
FT DOMAIN 567..774
FT /note="Peptidase C80"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01107"
FT REPEAT 1813..1832
FT /note="Cell wall-binding 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1833..1852
FT /note="Cell wall-binding 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1854..1873
FT /note="Cell wall-binding 3"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1876..1895
FT /note="Cell wall-binding 4"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1926..1945
FT /note="Cell wall-binding 5"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1946..1965
FT /note="Cell wall-binding 6"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1967..1986
FT /note="Cell wall-binding 7"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 1987..2006
FT /note="Cell wall-binding 8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2007..2026
FT /note="Cell wall-binding 9"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2057..2076
FT /note="Cell wall-binding 10"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2077..2097
FT /note="Cell wall-binding 11"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2099..2118
FT /note="Cell wall-binding 12"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2119..2138
FT /note="Cell wall-binding 13"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2139..2158
FT /note="Cell wall-binding 14"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2209..2224
FT /note="Cell wall-binding 15"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2227..2249
FT /note="Cell wall-binding 16"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2250..2269
FT /note="Cell wall-binding 17"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2270..2289
FT /note="Cell wall-binding 18"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2320..2339
FT /note="Cell wall-binding 19"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REPEAT 2340..2359
FT /note="Cell wall-binding 20"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00591"
FT REGION 1..91
FT /note="Four-helical bundle"
FT /evidence="ECO:0000269|PubMed:27023605"
FT REGION 96..468
FT /note="Glucosyltransferase region"
FT /evidence="ECO:0000303|PubMed:29146177"
FT REGION 544..799
FT /note="Autoprocessing region"
FT /evidence="ECO:0000303|PubMed:29146177"
FT REGION 800..1500
FT /note="Translocation region"
FT /evidence="ECO:0000303|PubMed:29146177"
FT REGION 1433..1438
FT /note="Interaction with host SEMA6A and SEMA6B"
FT /evidence="ECO:0000250|UniProtKB:P0DUB4"
FT REGION 1466..1471
FT /note="Interaction with host SEMA6A and SEMA6B"
FT /evidence="ECO:0000250|UniProtKB:P0DUB4"
FT REGION 1484..1495
FT /note="Interaction with host SEMA6A and SEMA6B"
FT /evidence="ECO:0000250|UniProtKB:P0DUB4"
FT REGION 1504..1511
FT /note="Interaction with host SEMA6A and SEMA6B"
FT /evidence="ECO:0000250|UniProtKB:P0DUB4"
FT REGION 1596..1601
FT /note="Interaction with host SEMA6A and SEMA6B"
FT /evidence="ECO:0000250|UniProtKB:P0DUB4"
FT REGION 1835..2364
FT /note="Receptor-binding (CROPS) region"
FT /evidence="ECO:0000303|PubMed:29146177"
FT ACT_SITE 653
FT /note="For protease activity"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01107"
FT ACT_SITE 698
FT /note="Nucleophile; for protease activity"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01107,
FT ECO:0000305|PubMed:27303685"
FT BINDING 101..103
FT /ligand="UDP-alpha-D-glucose"
FT /ligand_id="ChEBI:CHEBI:58885"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0000269|PubMed:18505687, ECO:0007744|PDB:2VKD,
FT ECO:0007744|PDB:2VKH, ECO:0007744|PDB:2VL8"
FT BINDING 139
FT /ligand="UDP-alpha-D-glucose"
FT /ligand_id="ChEBI:CHEBI:58885"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0000269|PubMed:18505687, ECO:0007744|PDB:2VKD,
FT ECO:0007744|PDB:2VKH, ECO:0007744|PDB:2VL8"
FT BINDING 265..270
FT /ligand="UDP-alpha-D-glucose"
FT /ligand_id="ChEBI:CHEBI:58885"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0000269|PubMed:18505687, ECO:0007744|PDB:2VKD,
FT ECO:0007744|PDB:2VKH, ECO:0007744|PDB:2VL8"
FT BINDING 286..288
FT /ligand="UDP-alpha-D-glucose"
FT /ligand_id="ChEBI:CHEBI:58885"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0000269|PubMed:18505687, ECO:0007744|PDB:2VKD,
FT ECO:0007744|PDB:2VKH, ECO:0007744|PDB:2VL8"
FT BINDING 288
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0007744|PDB:2VKD, ECO:0007744|PDB:2VKH"
FT BINDING 515
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0007744|PDB:2VKD, ECO:0007744|PDB:2VKH"
FT BINDING 518..520
FT /ligand="UDP-alpha-D-glucose"
FT /ligand_id="ChEBI:CHEBI:58885"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0007744|PDB:2VKD, ECO:0007744|PDB:2VKH"
FT BINDING 518
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:18325534,
FT ECO:0000269|PubMed:18505687, ECO:0007744|PDB:2VKH,
FT ECO:0007744|PDB:2VL8"
FT BINDING 545
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 546
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P18177"
FT BINDING 577
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 600
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 647
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 653
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P18177"
FT BINDING 757
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P18177"
FT BINDING 764
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 775
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT BINDING 792
FT /ligand="1D-myo-inositol hexakisphosphate"
FT /ligand_id="ChEBI:CHEBI:58130"
FT /evidence="ECO:0000250|UniProtKB:P16154"
FT SITE 543..544
FT /note="Cleavage; by autolysis"
FT /evidence="ECO:0000250|UniProtKB:P18177"
FT MUTAGEN 10
FT /note="Q->A: Does not affect phospholipid-binding and
FT cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 11
FT /note="K->I: Slightly reduced phospholipid-binding and
FT impaired cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 14
FT /note="Y->A: Strongly reduced phospholipid-binding without
FT affecting cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 15
FT /note="V->S: Strongly reduced phospholipid-binding and
FT impaired cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 16
FT /note="K->I: Slightly reduced phospholipid-binding."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 17..18
FT /note="FR->NA: Impaired localization to host cell membrane,
FT leading to impaired cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27303685"
FT MUTAGEN 17
FT /note="F->K: Strongly reduced phospholipid-binding and
FT impaired cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 18
FT /note="R->P: Strongly reduced phospholipid-binding and
FT impaired cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 20
FT /note="Q->A: Slightly reduced phospholipid-binding without
FT affecting cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 38
FT /note="S->A: Does not affect phospholipid-binding and
FT cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 68
FT /note="R->A: Strongly reduced phospholipid-binding without
FT affecting cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 78
FT /note="Y->A: Does not affect phospholipid-binding and
FT cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27023605"
FT MUTAGEN 286..288
FT /note="DVD->NVN: Abolished glucosyltransferase activity
FT without affecting localization to the host cell membrane."
FT /evidence="ECO:0000269|PubMed:24919149,
FT ECO:0000269|PubMed:27023605, ECO:0000269|PubMed:27303685"
FT MUTAGEN 698
FT /note="C->A: Abolished autoprocessing, leading to impaired
FT cytotoxicity."
FT /evidence="ECO:0000269|PubMed:27303685"
FT HELIX 6..12
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 22..35
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 42..62
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 69..88
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 96..101
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 109..121
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 125..131
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 138..157
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 158..160
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 161..164
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 168..196
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 202..213
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 218..233
FT /evidence="ECO:0007829|PDB:2VKH"
FT TURN 234..236
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 237..239
FT /evidence="ECO:0007829|PDB:2VKH"
FT TURN 240..242
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 249..260
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 265..280
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 282..285
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 295..298
FT /evidence="ECO:0007829|PDB:2VKD"
FT HELIX 309..324
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 335..337
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 340..351
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 356..358
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 374..378
FT /evidence="ECO:0007829|PDB:2VKH"
FT STRAND 381..389
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 394..419
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 424..437
FT /evidence="ECO:0007829|PDB:2VKH"
FT TURN 441..443
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 444..450
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 453..455
FT /evidence="ECO:0007829|PDB:2VKH"
FT TURN 456..458
FT /evidence="ECO:0007829|PDB:2VL8"
FT STRAND 459..461
FT /evidence="ECO:0007829|PDB:2VL8"
FT HELIX 464..468
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 471..482
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 495..498
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 499..501
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 505..507
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 513..518
FT /evidence="ECO:0007829|PDB:2VKH"
FT HELIX 529..536
FT /evidence="ECO:0007829|PDB:2VKH"
FT TURN 537..539
FT /evidence="ECO:0007829|PDB:2VKH"
SQ SEQUENCE 2364 AA; 270580 MW; EAD8A4467A89BDBB CRC64;
MNLVNKAQLQ KMVYVKFRIQ EDEYVAILNA LEEYHNMSES SVVEKYLKLK DINNLTDNYL
NTYKKSGRNK ALKKFKEYLT MEVLELKNNS LTPVEKNLHF IWIGGQINDT AINYINQWKD
VNSDYTVKVF YDSNAFLINT LKKTIVESAT NNTLESFREN LNDPEFDYNK FYRKRMEIIY
DKQKHFIDYY KSQIEENPEF IIDNIIKTYL SNEYSKDLEA LNKYIEESLN KITANNGNDI
RNLEKFADED LVRLYNQELV ERWNLAAASD ILRISMLKED GGVYLDVDIL PGIQPDLFKS
INKPDSITNT SWEMIKLEAI MKYKEYIPGY TSKNFDMLDE EVQRSFESAL SSKSDKSEIF
LPLDDIKVSP LEVKIAFANN SVINQALISL KDSYCSDLVI NQIKNRYKIL NDNLNPSINE
GTDFNTTMKI FSDKLASISN EDNMMFMIKI TNYLKVGFAP DVRSTINLSG PGVYTGAYQD
LLMFKDNSTN IHLLEPELRN FEFPKTKISQ LTEQEITSLW SFNQARAKSQ FEEYKKGYFE
GALGEDDNLD FAQNTVLDKD YVSKKILSSM KTRNKEYIHY IVQLQGDKIS YEASCNLFSK
DPYSSILYQK NIEGSETAYY YYVADAEIKE IDKYRIPYQI SNKRNIKLTF IGHGKSEFNT
DTFANLDVDS LSSEIETILN LAKADISPKY IEINLLGCNM FSYSISAEET YPGKLLLKIK
DRVSELMPSI SQDSITVSAN QYEVRINEEG KREILDHSGK WINKEESIIK DISSKEYISF
NPKENKIIVK SKYLHELSTL LQEIRNNANS SDIDLEKKVM LTECEINVAS NIDRQIVEGR
IEEAKNLTSD SINYIKNEFK LIESISDSLY DLKHQNGLDD SHFISFEDIS KTENGFRIRF
INKETGNSIF IETEKEIFSE YATHISKEIS NIKDTIFDNV NGKLVKKVNL DAAHEVNTLN
SAFFIQSLIE YNTTKESLSN LSVAMKVQVY AQLFSTGLNT ITDASKVVEL VSTALDETID
LLPTLSEGLP IIATIIDGVS LGAAIKELSE TNDPLLRQEI EAKIGIMAVN LTAASTAIVT
SALGIASGFS ILLVPLAGIS AGIPSLVNNE LILQDKATKV IDYFKHISLA ETEGAFTLLD
DKIIMPQDDL VLSEIDFNNN SITLGKCEIW RAEGGSGHTL TDDIDHFFSS PSITYRKPWL
SIYDVLNIKK EKIDFSKDLM VLPNAPNRVF GYEMGWTPGF RSLDNDGTKL LDRIRDHYEG
QFYWRYFAFI ADALITKLKP RYEDTNVRIN LDGNTRSFIV PVITTEQIRK NLSYSFYGSG
GSYSLSLSPY NMNIDLNLVE NDTWVIDVDN VVKNITIESD EIQKGELIEN ILSKLNIEDN
KIILNNHTIN FYGDINESNR FISLTFSILE DINIIIEIDL VSKSYKILLS GNCMKLIENS
SDIQQKIDHI GFNGEHQKYI PYSYIDNETK YNGFIDYSKK EGLFTAEFSN ESIIRNIYMP
DSNNLFIYSS KDLKDIRIIN KGDVKLLIGN YFKDDMKVSL SFTIEDTNTI KLNGVYLDEN
GVAQILKFMN NAKSALNTSN SLMNFLESIN IKNIFYNNLD PNIEFILDTN FIISGSNSIG
QFELICDKDK NIQPYFINFK IKETSYTLYV GNRQNLIVEP SYHLDDSGNI SSTVINFSQK
YLYGIDRYVN KVIIAPNLYT DEINITPVYK PNYICPEVII LDANYINEKI NVNINDLSIR
YVWDNDGSDL ILIANSEEDN QPQVKIRFVN VFKSDTAADK LSFNFSDKQD VSVSKIISTF
SLAAYSDGFF DYEFGLVSLD NDYFYINSFG NMVSGLIYIN DSLYYFKPPK NNLITGFTTI
DGNKYYFDPT KSGAASIGEI TIDGKDYYFN KQGILQVGVI NTSDGLKYFA PAGTLDENLE
GESVNFIGKL NIDGKIYYFE DNYRAAVEWK LLDDETYYFN PKTGEALKGL HQIGDNKYYF
DDNGIMQTGF ITINDKVFYF NNDGVMQVGY IEVNGKYFYF GKNGERQLGV FNTPDGFKFF
GPKDDDLGTE EGELTLYNGI LNFNGKIYFF DISNTAVVGW GTLDDGSTYY FDDNRAEACI
GLTVINDCKY YFDDNGIRQL GFITINDNIF YFSESGKIEL GYQNINGNYF YIDESGLVLI
GVFDTPDGYK YFAPLNTVND NIYGQAVKYS GLVRVNEDVY YFGETYKIET GWIENETDKY
YFDPETKKAY KGINVVDDIK YYFDENGIMR TGLISFENNN YYFNEDGKMQ FGYLNIKDKM
FYFGKDGKMQ IGVFNTPDGF KYFAHQNTLD ENFEGESINY TGWLDLDGKR YYFTDEYIAA
TGSLTIDGYN YYFDPDTAEL VVSE
