TACY_STRP2
ID TACY_STRP2 Reviewed; 471 AA.
AC Q04IN8; P11990;
DT 06-MAR-2007, integrated into UniProtKB/Swiss-Prot.
DT 14-NOV-2006, sequence version 1.
DT 03-AUG-2022, entry version 94.
DE RecName: Full=Pneumolysin;
DE Short=PLY;
DE AltName: Full=Thiol-activated cytolysin;
GN Name=ply; OrderedLocusNames=SPD_1726;
OS Streptococcus pneumoniae serotype 2 (strain D39 / NCTC 7466).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=373153;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-17.
RC STRAIN=D39 / NCTC 7466;
RX PubMed=3552992; DOI=10.1128/iai.55.5.1184-1189.1987;
RA Walker J.A., Allen R.L., Falmagne P., Johnson M.K., Boulnois G.J.;
RT "Molecular cloning, characterization, and complete nucleotide sequence of
RT the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus
RT pneumoniae.";
RL Infect. Immun. 55:1184-1189(1987).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=D39 / NCTC 7466;
RX PubMed=17041037; DOI=10.1128/jb.01148-06;
RA Lanie J.A., Ng W.-L., Kazmierczak K.M., Andrzejewski T.M., Davidsen T.M.,
RA Wayne K.J., Tettelin H., Glass J.I., Winkler M.E.;
RT "Genome sequence of Avery's virulent serotype 2 strain D39 of Streptococcus
RT pneumoniae and comparison with that of unencapsulated laboratory strain
RT R6.";
RL J. Bacteriol. 189:38-51(2007).
RN [3]
RP FUNCTION, DOMAIN, AND MUTAGENESIS OF CYS-428; TRP-433; GLU-434; TRP-435 AND
RP TRP-436.
RC STRAIN=D39 / NCTC 7466;
RX PubMed=9445384; DOI=10.1042/bj3290571;
RA Korchev Y.E., Bashford C.L., Pederzolli C., Pasternak C.A., Morgan P.J.,
RA Andrew P.W., Mitchell T.J.;
RT "A conserved tryptophan in pneumolysin is a determinant of the
RT characteristics of channels formed by pneumolysin in cells and planar lipid
RT bilayers.";
RL Biochem. J. 329:571-577(1998).
RN [4]
RP FUNCTION, SUBUNIT, SUBCELLULAR LOCATION, AND DOMAIN.
RX PubMed=15851031; DOI=10.1016/j.cell.2005.02.033;
RA Tilley S.J., Orlova E.V., Gilbert R.J., Andrew P.W., Saibil H.R.;
RT "Structural basis of pore formation by the bacterial toxin pneumolysin.";
RL Cell 121:247-256(2005).
RN [5]
RP FUNCTION, CHOLESTEROL-BINDING, AND MUTAGENESIS OF 459-THR-LEU-460.
RX PubMed=20145114; DOI=10.1073/pnas.0911581107;
RA Farrand A.J., LaChapelle S., Hotze E.M., Johnson A.E., Tweten R.K.;
RT "Only two amino acids are essential for cytolytic toxin recognition of
RT cholesterol at the membrane surface.";
RL Proc. Natl. Acad. Sci. U.S.A. 107:4341-4346(2010).
RN [6] {ECO:0007744|PDB:5CR6, ECO:0007744|PDB:5CR8}
RP X-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS), FUNCTION, ACTIVITY REGULATION,
RP DOMAIN, AND MUTAGENESIS OF THR-88; ARG-147; ASP-205; ARG-226; THR-304;
RP ASN-339 AND 434-GLU--ARG-437.
RC STRAIN=D39 / NCTC 7466;
RX PubMed=26333773; DOI=10.1038/srep13293;
RA Marshall J.E., Faraj B.H., Gingras A.R., Lonnen R., Sheikh M.A.,
RA El-Mezgueldi M., Moody P.C., Andrew P.W., Wallis R.;
RT "The Crystal Structure of Pneumolysin at 2.0 A Resolution Reveals the
RT Molecular Packing of the Pre-pore Complex.";
RL Sci. Rep. 5:13293-13293(2015).
RN [7] {ECO:0007744|PDB:4ZGH}
RP X-RAY CRYSTALLOGRAPHY (2.90 ANGSTROMS) OF 2-470, DOMAIN, AND MUTAGENESIS OF
RP LYS-18; 66-ASN--SER-68; ASN-66; SER-68 AND LYS-208.
RC STRAIN=D39 / NCTC 7466;
RX PubMed=26403197; DOI=10.1038/srep14352;
RA Lawrence S.L., Feil S.C., Morton C.J., Farrand A.J., Mulhern T.D.,
RA Gorman M.A., Wade K.R., Tweten R.K., Parker M.W.;
RT "Crystal structure of Streptococcus pneumoniae pneumolysin provides key
RT insights into early steps of pore formation.";
RL Sci. Rep. 5:14352-14352(2015).
RN [8] {ECO:0007744|PDB:5AOD}
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS), DOMAIN, AND MUTAGENESIS OF THR-63;
RP ASN-85; ASP-102; LYS-152; GLN-191; ASP-205; GLU-260; GLU-264 AND THR-405.
RX PubMed=27796097; DOI=10.1021/acs.nanolett.6b04219;
RA van Pee K., Mulvihill E., Mueller D.J., Yildiz O.;
RT "Unraveling the Pore-Forming Steps of Pneumolysin from Streptococcus
RT pneumoniae.";
RL Nano Lett. 16:7915-7924(2016).
RN [9] {ECO:0007744|PDB:5AOE, ECO:0007744|PDB:5AOF, ECO:0007744|PDB:5LY6}
RP STRUCTURE BY ELECTRON MICROSCOPY (4.50 ANGSTROMS) OF WHOLE PORE, X-RAY
RP CRYSTALLOGRAPHY (2.45 ANGSTROMS) OF NON-HEMOLYTIC MUTANTS, PORE FORMATION,
RP SUBUNIT, DOMAIN, AND MUTAGENESIS OF 146-ALA-ARG-147 AND ASP-168.
RC STRAIN=D39 / NCTC 7466;
RX PubMed=28323617; DOI=10.7554/elife.23644;
RA van Pee K., Neuhaus A., D'Imprima E., Mills D.J., Kuhlbrandt W., Yildiz O.;
RT "CryoEM structures of membrane pore and prepore complex reveal cytolytic
RT mechanism of Pneumolysin.";
RL Elife 6:0-0(2017).
CC -!- FUNCTION: A cholesterol-dependent toxin that causes cytolysis by
CC forming pores in cholesterol-containing host membranes. After binding
CC to target membranes, the protein undergoes a major conformation change,
CC leading to its insertion in the host membrane and formation of an
CC oligomeric pore complex. Cholesterol is required for binding to host
CC membranes, membrane insertion and pore formation; cholesterol binding
CC is mediated by a Thr-Leu pair in the C-terminus. Can be reversibly
CC inactivated by oxidation. {ECO:0000269|PubMed:15851031,
CC ECO:0000269|PubMed:20145114, ECO:0000269|PubMed:26333773,
CC ECO:0000269|PubMed:28323617, ECO:0000269|PubMed:9445384}.
CC -!- ACTIVITY REGULATION: Erythrocytes hemolysis is inhibited by
CC cholesterol. {ECO:0000269|PubMed:26333773}.
CC -!- SUBUNIT: Elongated monomers align along their lengths, indicating
CC intersubunit contacts and suggesting the prepore structure
CC (PubMed:15851031, PubMed:26333773, PubMed:26403197, PubMed:27796097,
CC PubMed:28323617). Modeling based on cryo-EM shows a homooligomeric pore
CC complex containing 38-44 subunits; when inserted in the host membrane
CC (Probable). The size of isolated pores is detergent-dependent; in
CC amphipol A8-35 homogenous rings form with 42 subunits
CC (PubMed:28323617). {ECO:0000269|PubMed:15851031,
CC ECO:0000269|PubMed:26333773, ECO:0000269|PubMed:26403197,
CC ECO:0000269|PubMed:27796097, ECO:0000269|PubMed:28323617,
CC ECO:0000305|PubMed:15851031}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:P0C2J9}. Host
CC cell membrane {ECO:0000269|PubMed:28323617,
CC ECO:0000305|PubMed:15851031}; Multi-pass membrane protein
CC {ECO:0000269|PubMed:28323617}. Note=Secreted as soluble protein by the
CC accessory Sec system (By similarity). It then inserts into the host
CC cell membrane and forms pores formed by transmembrane beta-strands
CC (PubMed:15851031, PubMed:28323617). {ECO:0000250|UniProtKB:P0C2J9,
CC ECO:0000269|PubMed:15851031, ECO:0000269|PubMed:28323617}.
CC -!- DOMAIN: A highly conserved undecapeptide in the C-terminus has residues
CC important for membrane binding and cell lysis (PubMed:9445384). Mature
CC protein has 3 discontinuous domains; D1, D2, D3 followed by C-terminal
CC D4 (PubMed:15851031, PubMed:26333773, PubMed:26403197, PubMed:27796097,
CC PubMed:28323617). The domains rearrange substantially upon membrane
CC insertion, in particular alpha-helices in D3 refold to form the
CC transmembrane beta-strands (PubMed:15851031, PubMed:28323617). The
CC relative position of domain D4 changes, allowing it to interact with
CC host membranes (PubMed:26333773, PubMed:27796097, PubMed:28323617).
CC {ECO:0000269|PubMed:15851031, ECO:0000269|PubMed:26333773,
CC ECO:0000269|PubMed:26403197, ECO:0000269|PubMed:27796097,
CC ECO:0000269|PubMed:28323617, ECO:0000269|PubMed:9445384}.
CC -!- SIMILARITY: Belongs to the cholesterol-dependent cytolysin family.
CC {ECO:0000305}.
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DR EMBL; X52474; CAA36714.1; -; Genomic_DNA.
DR EMBL; M17717; AAA26915.1; -; Genomic_DNA.
DR EMBL; CP000410; ABJ53672.1; -; Genomic_DNA.
DR RefSeq; WP_001284359.1; NC_008533.2.
DR PDB; 4ZGH; X-ray; 2.90 A; A=2-470.
DR PDB; 5AOD; X-ray; 2.40 A; A=1-471.
DR PDB; 5AOE; X-ray; 2.50 A; A/B=1-471.
DR PDB; 5AOF; X-ray; 2.45 A; A=1-470.
DR PDB; 5CR6; X-ray; 1.98 A; D=1-471.
DR PDB; 5CR8; X-ray; 2.05 A; A/D=359-471.
DR PDB; 5LY6; EM; 4.50 A; B=1-471.
DR PDBsum; 4ZGH; -.
DR PDBsum; 5AOD; -.
DR PDBsum; 5AOE; -.
DR PDBsum; 5AOF; -.
DR PDBsum; 5CR6; -.
DR PDBsum; 5CR8; -.
DR PDBsum; 5LY6; -.
DR AlphaFoldDB; Q04IN8; -.
DR SMR; Q04IN8; -.
DR STRING; 373153.SPD_1726; -.
DR EnsemblBacteria; ABJ53672; ABJ53672; SPD_1726.
DR GeneID; 60232858; -.
DR GeneID; 66806991; -.
DR KEGG; spd:SPD_1726; -.
DR eggNOG; ENOG502Z7ST; Bacteria.
DR HOGENOM; CLU_026912_1_0_9; -.
DR OMA; NDRTYPG; -.
DR OrthoDB; 1219984at2; -.
DR BioCyc; SPNE373153:G1G6V-1864-MON; -.
DR PHI-base; PHI:6282; -.
DR PHI-base; PHI:7994; -.
DR Proteomes; UP000001452; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0020002; C:host cell plasma membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0015485; F:cholesterol binding; IEA:InterPro.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0044179; P:hemolysis in another organism; IEA:UniProtKB-KW.
DR Gene3D; 2.60.40.1430; -; 1.
DR Gene3D; 3.90.840.10; -; 1.
DR InterPro; IPR035390; Thiol_cytolys_C.
DR InterPro; IPR038700; Thiol_cytolys_C_sf.
DR InterPro; IPR001869; Thiol_cytolysin.
DR InterPro; IPR036363; Thiol_cytolysin_ab_sf.
DR InterPro; IPR036359; Thiol_cytolysin_sf.
DR Pfam; PF17440; Thiol_cytolys_C; 1.
DR Pfam; PF01289; Thiol_cytolysin; 1.
DR PRINTS; PR01400; TACYTOLYSIN.
DR SUPFAM; SSF56978; SSF56978; 1.
DR PROSITE; PS00481; THIOL_CYTOLYSINS; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytolysis; Direct protein sequencing; Hemolysis;
KW Host cell membrane; Host membrane; Lipid-binding; Membrane; Secreted;
KW Toxin; Transmembrane; Transmembrane beta strand; Virulence.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:3552992"
FT CHAIN 2..471
FT /note="Pneumolysin"
FT /id="PRO_0000279546"
FT TRANSMEM 158..171
FT /note="Beta stranded"
FT /evidence="ECO:0000269|PubMed:28323617,
FT ECO:0007744|PDB:5LY6"
FT TRANSMEM 178..187
FT /note="Beta stranded"
FT /evidence="ECO:0000269|PubMed:28323617,
FT ECO:0007744|PDB:5LY6"
FT TRANSMEM 256..265
FT /note="Beta stranded"
FT /evidence="ECO:0000269|PubMed:28323617,
FT ECO:0007744|PDB:5LY6"
FT TRANSMEM 273..285
FT /note="Beta stranded"
FT /evidence="ECO:0000269|PubMed:28323617,
FT ECO:0007744|PDB:5LY6"
FT MOTIF 427..437
FT /note="Conserved undecapeptide"
FT /evidence="ECO:0000305"
FT MOTIF 459..460
FT /note="Cholesterol binding"
FT /evidence="ECO:0000269|PubMed:20145114"
FT MUTAGEN 18
FT /note="K->A: Loss of hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26403197"
FT MUTAGEN 63
FT /note="T->A: 15% hemolytic activity, forms incomplete rings
FT on liposomes."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 66..68
FT /note="NDS->WDW: Loss of hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26403197"
FT MUTAGEN 66
FT /note="N->W: 50% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26403197"
FT MUTAGEN 68
FT /note="S->W: Wild-type hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26403197"
FT MUTAGEN 85
FT /note="N->A: 44% hemolytic activity, forms incomplete rings
FT on liposomes."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 88
FT /note="T->E: 4% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 102
FT /note="D->A: 1% hemolytic activity, forms linear oligomers
FT on liposomes rather than pores."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 146..147
FT /note="Missing: Loss of hemolytic activity, does not bind
FT membrane, does not form pores."
FT /evidence="ECO:0000269|PubMed:28323617"
FT MUTAGEN 147
FT /note="R->E: 2% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 152
FT /note="K->D: 12% hemolytic activity; when associated with
FT K-260."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 168
FT /note="D->A: 20% hemolytic activity, inhibits membrane
FT insertion, forms prepores on liposomes which detach easily
FT and rarely convert to pores."
FT /evidence="ECO:0000269|PubMed:28323617"
FT MUTAGEN 191
FT /note="Q->E: 42% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 205
FT /note="D->A: 17% hemolytic activity, forms linear oligomers
FT on liposomes rather than pores."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 205
FT /note="D->R: Loss of hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 208
FT /note="K->A: 4% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26403197"
FT MUTAGEN 226
FT /note="R->A: 6% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 260
FT /note="E->K: 22% hemolytic activity. 12% hemolytic
FT activity; when associated with D-152."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 264
FT /note="E->D: 76% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 304
FT /note="T->R: 320-fold decrease in hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 339
FT /note="N->R: Loss of hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 405
FT /note="T->L: 15% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:27796097"
FT MUTAGEN 428
FT /note="C->A: Wild-type hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 428
FT /note="C->G: 3% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 428
FT /note="C->S: 25% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 433
FT /note="W->F: 1% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 434..437
FT /note="EWWR->AWWA: 5% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:26333773"
FT MUTAGEN 434
FT /note="E->D: 50% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 434
FT /note="E->Q: 20% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 435
FT /note="W->F: 20% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 436
FT /note="W->F: 50% hemolytic activity."
FT /evidence="ECO:0000269|PubMed:9445384"
FT MUTAGEN 459..460
FT /note="TL->GG: Loss of hemolytic activity, loss of host
FT membrane binding, loss of cholesterol binding."
FT /evidence="ECO:0000269|PubMed:20145114"
FT HELIX 2..12
FT /evidence="ECO:0007829|PDB:5CR6"
FT TURN 17..21
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 22..24
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 33..39
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 42..61
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 75..79
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 80..83
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 97..101
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 108..111
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 112..117
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 120..137
FT /evidence="ECO:0007829|PDB:5CR6"
FT TURN 138..141
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 147..154
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 158..165
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 170..176
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 180..184
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 189..203
FT /evidence="ECO:0007829|PDB:5CR6"
FT TURN 210..213
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 222..226
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 229..232
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 234..252
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 259..267
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 277..283
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 285..292
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 301..305
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 307..309
FT /evidence="ECO:0007829|PDB:5CR6"
FT HELIX 310..315
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 322..324
FT /evidence="ECO:0007829|PDB:4ZGH"
FT STRAND 327..339
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 344..359
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 361..367
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 369..371
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 373..384
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 390..396
FT /evidence="ECO:0007829|PDB:5CR6"
FT TURN 398..401
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 406..414
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 418..431
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 434..444
FT /evidence="ECO:0007829|PDB:5CR6"
FT STRAND 449..469
FT /evidence="ECO:0007829|PDB:5CR6"
SQ SEQUENCE 471 AA; 52899 MW; 966AF46FE2EA5792 CRC64;
MANKAVNDFI LAMNYDKKKL LTHQGESIEN RFIKEGNQLP DEFVVIERKK RSLSTNTSDI
SVTATNDSRL YPGALLVVDE TLLENNPTLL AVDRAPMTYS IDLPGLASSD SFLQVEDPSN
SSVRGAVNDL LAKWHQDYGQ VNNVPARMQY EKITAHSMEQ LKVKFGSDFE KTGNSLDIDF
NSVHSGEKQI QIVNFKQIYY TVSVDAVKNP GDVFQDTVTV EDLKQRGISA ERPLVYISSV
AYGRQVYLKL ETTSKSDEVE AAFEALIKGV KVAPQTEWKQ ILDNTEVKAV ILGGDPSSGA
RVVTGKVDMV EDLIQEGSRF TADHPGLPIS YTTSFLRDNV VATFQNSTDY VETKVTAYRN
GDLLLDHSGA YVAQYYITWD ELSYDHQGKE VLTPKAWDRN GQDLTAHFTT SIPLKGNVRN
LSVKIRECTG LAWEWWRTVY EKTDLPLVRK RTISIWGTTL YPQVEDKVEN D
