HLYC_ECOLX
ID HLYC_ECOLX Reviewed; 170 AA.
AC P06736;
DT 01-JAN-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1988, sequence version 1.
DT 03-AUG-2022, entry version 87.
DE RecName: Full=Protein-lysine myristoyltransferase HlyC {ECO:0000305};
DE EC=2.3.1.- {ECO:0000269|PubMed:11695909, ECO:0000269|PubMed:32461253};
DE AltName: Full=Hemolysin C;
DE AltName: Full=Hemolysin-activating lysine-acyltransferase HlyC;
DE AltName: Full=Toxin-activating protein C, PHLY152;
GN Name=hlyC {ECO:0000303|PubMed:9521785};
OS Escherichia coli.
OG Plasmid IncI2 pHLY152.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Hess J., Wels W., Vogel M., Goebel W.;
RT "Nucleotide sequence of a plasmid-encoded hemolysin determinant and its
RT comparison with a corresponding chromosomal hemolysin sequence.";
RL FEMS Microbiol. Lett. 34:1-11(1986).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=6394952; DOI=10.1007/bf00330963;
RA Juarez A., Hughes C., Vogel M., Goebel W.;
RT "Expression and regulation of the plasmid-encoded hemolysin determinant of
RT Escherichia coli.";
RL Mol. Gen. Genet. 197:196-203(1984).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2990440; DOI=10.1007/978-1-4613-2447-8_55;
RA Goebel W., Hacker J., Knapp S., Then I., Wagner W., Hughes C., Juarez A.;
RT "Structure, function, and regulation of the plasmid-encoded hemolysin
RT determinant of Escherichia coli.";
RL Basic Life Sci. 30:791-805(1985).
RN [4]
RP FUNCTION, AND SUBUNIT.
RX PubMed=9521785; DOI=10.1021/bi971588y;
RA Trent M.S., Worsham L.M., Ernst-Fonberg M.L.;
RT "The biochemistry of hemolysin toxin activation: characterization of HlyC,
RT an internal protein acyltransferase.";
RL Biochemistry 37:4644-4652(1998).
RN [5]
RP FUNCTION, ACTIVE SITE, AND MUTAGENESIS OF SER-20; HIS-23; CYS-57; SER-58
RP AND SER-76.
RX PubMed=10079090; DOI=10.1021/bi982491u;
RA Trent M.S., Worsham L.M., Ernst-Fonberg M.L.;
RT "HlyC, the internal protein acyltransferase that activates hemolysin toxin:
RT role of conserved histidine, serine, and cysteine residues in enzymatic
RT activity as probed by chemical modification and site-directed
RT mutagenesis.";
RL Biochemistry 38:3433-3439(1999).
RN [6]
RP MUTAGENESIS OF ARG-24; TYR-70; ARG-87 AND TYR-150.
RX PubMed=10393560; DOI=10.1021/bi990138y;
RA Trent M.S., Worsham L.M., Ernst-Fonberg M.L.;
RT "HlyC, the internal protein acyltransferase that activates hemolysin toxin:
RT the role of conserved tyrosine and arginine residues in enzymatic activity
RT as probed by chemical modification and site-directed mutagenesis.";
RL Biochemistry 38:8831-8838(1999).
RN [7]
RP FUNCTION, AND MUTAGENESIS OF GLY-11; PRO-53; GLY-85; ASP-86 AND PRO-96.
RX PubMed=10413532; DOI=10.1021/bi9905617;
RA Trent M.S., Worsham L.M., Ernst-Fonberg M.L.;
RT "HlyC, the internal protein acyltransferase that activates hemolysin toxin:
RT roles of various conserved residues in enzymatic activity as probed by
RT site-directed mutagenesis.";
RL Biochemistry 38:9541-9548(1999).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF 19-SER-SER-20; SER-20;
RP HIS-23; GLN-43 AND CYS-57.
RX PubMed=11695909; DOI=10.1021/bi011032h;
RA Worsham L.M., Trent M.S., Earls L., Jolly C., Ernst-Fonberg M.L.;
RT "Insights into the catalytic mechanism of HlyC, the internal protein
RT acyltransferase that activates Escherichia coli hemolysin toxin.";
RL Biochemistry 40:13607-13616(2001).
RN [9]
RP FUNCTION, DEGRADATION, AND MUTAGENESIS OF 128-GLY--VAL-130;
RP 128-GLY-LYS-129; GLY-128 AND LYS-129.
RX PubMed=11278516; DOI=10.1074/jbc.m009514200;
RA Guzman-Verri C., Chaves-Olarte E., Garcia F., Arvidson S., Moreno E.;
RT "In vivo proteolytic degradation of the Escherichia coli acyltransferase
RT HlyC.";
RL J. Biol. Chem. 276:16660-16666(2001).
RN [10]
RP FUNCTION.
RX PubMed=15065878; DOI=10.1021/bi035919k;
RA Langston K.G., Worsham L.M., Earls L., Ernst-Fonberg M.L.;
RT "Activation of hemolysin toxin: relationship between two internal protein
RT sites of acylation.";
RL Biochemistry 43:4338-4346(2004).
RN [11]
RP FUNCTION, ACTIVITY REGULATION, HEME-BINDING, AND MUTAGENESIS OF
RP 151-HIS-HIS-152.
RX PubMed=29908817; DOI=10.1016/j.bbagen.2018.06.012;
RA Peherstorfer S., Brewitz H.H., Paul George A.A., Wissbrock A., Adam J.M.,
RA Schmitt L., Imhof D.;
RT "Insights into mechanism and functional consequences of heme binding to
RT hemolysin-activating lysine acyltransferase HlyC from Escherichia coli.";
RL Biochim. Biophys. Acta 1862:1964-1972(2018).
RN [12]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=32461253; DOI=10.1074/jbc.ra120.014122;
RA Osickova A., Khaliq H., Masin J., Jurnecka D., Sukova A., Fiser R.,
RA Holubova J., Stanek O., Sebo P., Osicka R.;
RT "Acyltransferase-mediated selection of the length of the fatty acyl chain
RT and of the acylation site governs activation of bacterial RTX toxins.";
RL J. Biol. Chem. 295:9268-9280(2020).
CC -!- FUNCTION: Protein-lysine myristoyltransferase that catalyzes
CC myristoylation of the protoxin (HlyA) at two internal lysine residues,
CC thereby converting it to the active toxin.
CC {ECO:0000269|PubMed:10079090, ECO:0000269|PubMed:10413532,
CC ECO:0000269|PubMed:11278516, ECO:0000269|PubMed:11695909,
CC ECO:0000269|PubMed:15065878, ECO:0000269|PubMed:29908817,
CC ECO:0000269|PubMed:32461253, ECO:0000269|PubMed:9521785}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-lysyl-[protein] + tetradecanoyl-[ACP] = H(+) + holo-[ACP] +
CC N(6)-tetradecanoyl-L-lysyl-[protein]; Xref=Rhea:RHEA:70611,
CC Rhea:RHEA-COMP:9648, Rhea:RHEA-COMP:9685, Rhea:RHEA-COMP:9752,
CC Rhea:RHEA-COMP:15437, ChEBI:CHEBI:15378, ChEBI:CHEBI:29969,
CC ChEBI:CHEBI:64479, ChEBI:CHEBI:78477, ChEBI:CHEBI:141129;
CC Evidence={ECO:0000269|PubMed:11695909, ECO:0000269|PubMed:32461253};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70612;
CC Evidence={ECO:0000269|PubMed:11695909, ECO:0000269|PubMed:32461253};
CC -!- ACTIVITY REGULATION: The acyltransferase activity is inhibited by heme.
CC {ECO:0000269|PubMed:29908817}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.6 uM for tetradecanoyl-CoA {ECO:0000269|PubMed:11695909};
CC KM=3.6 uM for HlyA peptide {ECO:0000269|PubMed:11695909};
CC Vmax=4667 pmol/min/mg enzyme {ECO:0000269|PubMed:11695909};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:9521785}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- PTM: Proteolytically cleaved by the protease systems ClpAP, ClpXP and
CC FtsH, leading to its degradation. {ECO:0000269|PubMed:11278516}.
CC -!- SIMILARITY: Belongs to the RTX toxin acyltransferase family.
CC {ECO:0000305}.
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DR EMBL; M14107; AAA98232.1; -; Genomic_DNA.
DR EMBL; X01072; CAA25535.1; -; Genomic_DNA.
DR EMBL; M35668; AAA23980.1; -; Genomic_DNA.
DR PIR; I41288; I41288.
DR RefSeq; WP_001372478.1; NZ_SVID01000104.1.
DR AlphaFoldDB; P06736; -.
DR SMR; P06736; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0018030; F:peptidyl-lysine N6-myristoyltransferase activity; IDA:UniProtKB.
DR GO; GO:0044179; P:hemolysis in another organism; IEA:UniProtKB-KW.
DR GO; GO:0009404; P:toxin metabolic process; IEA:InterPro.
DR InterPro; IPR003996; RTX_toxin-activating_protC_bac.
DR Pfam; PF02794; HlyC; 1.
DR PRINTS; PR01489; RTXTOXINC.
PE 1: Evidence at protein level;
KW Acyltransferase; Cytolysis; Cytoplasm; Heme; Hemolysis; Iron;
KW Metal-binding; Plasmid; Transferase; Virulence.
FT CHAIN 1..170
FT /note="Protein-lysine myristoyltransferase HlyC"
FT /id="PRO_0000217875"
FT ACT_SITE 23
FT /evidence="ECO:0000305|PubMed:10079090"
FT ACT_SITE 92
FT /evidence="ECO:0000250|UniProtKB:P55132"
FT BINDING 151
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000305|PubMed:29908817"
FT MUTAGEN 11
FT /note="G->A: Strongly decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10413532"
FT MUTAGEN 19..20
FT /note="SS->AA: Significant loss of acyltransferase
FT activity."
FT /evidence="ECO:0000269|PubMed:11695909"
FT MUTAGEN 20
FT /note="S->A,C,T,H: Significant loss of acyltransferase
FT activity."
FT /evidence="ECO:0000269|PubMed:10079090,
FT ECO:0000269|PubMed:11695909"
FT MUTAGEN 23
FT /note="H->A,C,S,D,K,Y: Complete loss of acyltransferase
FT activity."
FT /evidence="ECO:0000269|PubMed:10079090,
FT ECO:0000269|PubMed:11695909"
FT MUTAGEN 24
FT /note="R->A,K: Little effect."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 43
FT /note="Q->A: Decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:11695909"
FT MUTAGEN 53
FT /note="P->A: Strongly decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10413532"
FT MUTAGEN 57
FT /note="C->A: Does not affect acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10079090,
FT ECO:0000269|PubMed:11695909"
FT MUTAGEN 58
FT /note="S->A: Little effect."
FT /evidence="ECO:0000269|PubMed:10079090"
FT MUTAGEN 70
FT /note="Y->F: No effect."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 70
FT /note="Y->G: Strongly decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 76
FT /note="S->A: Slightly decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10079090"
FT MUTAGEN 85
FT /note="G->A: Abolished acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10413532"
FT MUTAGEN 86
FT /note="D->A: Decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10413532"
FT MUTAGEN 87
FT /note="R->A,K: Little effect."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 96
FT /note="P->A: Strongly decreased acyltransferase activity."
FT /evidence="ECO:0000269|PubMed:10413532"
FT MUTAGEN 128..130
FT /note="Missing: Abolished ability to activate HlyA. Does
FT not affect protein stability."
FT /evidence="ECO:0000269|PubMed:11278516"
FT MUTAGEN 128..129
FT /note="GK->VI: Abolished ability to activate HlyA. Does not
FT affect protein stability."
FT /evidence="ECO:0000269|PubMed:11278516"
FT MUTAGEN 128
FT /note="G->V: Abolished ability to activate HlyA. Does not
FT affect protein stability."
FT /evidence="ECO:0000269|PubMed:11278516"
FT MUTAGEN 129
FT /note="K->I: Reduced ability to activate HlyA."
FT /evidence="ECO:0000269|PubMed:11278516"
FT MUTAGEN 150
FT /note="Y->F: No effect."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 150
FT /note="Y->G: Great loss of activity."
FT /evidence="ECO:0000269|PubMed:10393560"
FT MUTAGEN 151..152
FT /note="HH->AA: Abolished heme-binding."
FT /evidence="ECO:0000269|PubMed:29908817"
SQ SEQUENCE 170 AA; 19770 MW; 754DB786609734EF CRC64;
MNINKPLEIL GHVSWLWASS PLHRNWPVSL FAINVLPAIQ ANQYVLLTRD DYPVAYCSWA
NLSLENEIKY LNDVTSLVAE DWTSGDRKWF IDWIAPFGDN GALYKYMRKK FPDELFRAIR
VDPKTHVGKV SEFHGGKIDK QLANKIFKQY HHELITEVKR KSDFNFSLTG
