LEP4_PSEAE
ID LEP4_PSEAE Reviewed; 290 AA.
AC P22610;
DT 01-AUG-1991, integrated into UniProtKB/Swiss-Prot.
DT 08-DEC-2000, sequence version 3.
DT 03-AUG-2022, entry version 140.
DE RecName: Full=Prepilin leader peptidase/N-methyltransferase {ECO:0000303|PubMed:8340405};
DE AltName: Full=Protein PilD;
DE AltName: Full=Protein secretion protein XCPA;
DE Includes:
DE RecName: Full=Leader peptidase;
DE EC=3.4.23.43 {ECO:0000269|PubMed:23255525, ECO:0000269|PubMed:8096341, ECO:0000269|PubMed:8340405};
DE AltName: Full=Prepilin peptidase;
DE Includes:
DE RecName: Full=N-methyltransferase;
DE EC=2.1.1.- {ECO:0000269|PubMed:23255525, ECO:0000269|PubMed:8096341, ECO:0000269|PubMed:8340405};
GN Name=pilD; Synonyms=xcpA; OrderedLocusNames=PA4528;
OS Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM
OS 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=208964;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=PAK;
RX PubMed=1971619; DOI=10.1128/jb.172.6.2911-2919.1990;
RA Nunn D., Bergman S., Lory S.;
RT "Products of three accessory genes, pilB, pilC, and pilD, are required for
RT biogenesis of Pseudomonas aeruginosa pili.";
RL J. Bacteriol. 172:2911-2919(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=1898929; DOI=10.1128/jb.173.2.479-486.1991;
RA Bally M., Ball G., Badere A., Lazdunski A.;
RT "Protein secretion in Pseudomonas aeruginosa: the xcpA gene encodes an
RT integral inner membrane protein homologous to Klebsiella pneumoniae
RT secretion function protein PulO.";
RL J. Bacteriol. 173:479-486(1991).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=10984043; DOI=10.1038/35023079;
RA Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J.,
RA Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT "Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic
RT pathogen.";
RL Nature 406:959-964(2000).
RN [4]
RP REVIEW.
RX PubMed=9224881; DOI=10.1016/s0378-1119(96)00830-x;
RA Lory S., Strom M.S.;
RT "Structure-function relationship of type-IV prepilin peptidase of
RT Pseudomonas aeruginosa -- a review.";
RL Gene 192:117-121(1997).
RN [5]
RP REVIEW.
RX PubMed=8057924; DOI=10.1016/0076-6879(94)35168-6;
RA Strom M.S., Nunn D.N., Lory S.;
RT "Posttranslational processing of type IV prepilin and homologs by PilD of
RT Pseudomonas aeruginosa.";
RL Methods Enzymol. 235:527-540(1994).
RN [6]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8096341; DOI=10.1073/pnas.90.6.2404;
RA Strom M.S., Nunn D.N., Lory S.;
RT "A single bifunctional enzyme, PilD, catalyzes cleavage and N-methylation
RT of proteins belonging to the type IV pilin family.";
RL Proc. Natl. Acad. Sci. U.S.A. 90:2404-2408(1993).
RN [7]
RP FUNCTION, AND SUBCELLULAR LOCATION.
RC STRAIN=PAK;
RX PubMed=8331069; DOI=10.1128/jb.175.14.4375-4382.1993;
RA Nunn D.N., Lory S.;
RT "Cleavage, methylation, and localization of the Pseudomonas aeruginosa
RT export proteins XcpT, -U, -V, and -W.";
RL J. Bacteriol. 175:4375-4382(1993).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF CYS-72; CYS-75; CYS-97 AND
RP CYS-100.
RX PubMed=8340405; DOI=10.1016/s0021-9258(18)82324-9;
RA Strom M.S., Bergman P., Lory S.;
RT "Identification of active-site cysteines in the conserved domain of PilD,
RT the bifunctional type IV pilin leader peptidase/N-methyltransferase of
RT Pseudomonas aeruginosa.";
RL J. Biol. Chem. 268:15788-15794(1993).
RN [9]
RP MUTAGENESIS OF GLY-95 AND LYS-96.
RX PubMed=9668097; DOI=10.1074/jbc.273.30.19120;
RA Pepe J.C., Lory S.;
RT "Amino acid substitutions in PilD, a bifunctional enzyme of Pseudomonas
RT aeruginosa. Effect on leader peptidase and N-methyltransferase activities
RT in vitro and in vivo.";
RL J. Biol. Chem. 273:19120-19129(1998).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, AND COFACTOR.
RX PubMed=23255525; DOI=10.1002/mbo3.51;
RA Aly K.A., Beebe E.T., Chan C.H., Goren M.A., Sepulveda C., Makino S.,
RA Fox B.G., Forest K.T.;
RT "Cell-free production of integral membrane aspartic acid proteases reveals
RT zinc-dependent methyltransferase activity of the Pseudomonas aeruginosa
RT prepilin peptidase PilD.";
RL MicrobiologyOpen 2:94-104(2013).
CC -!- FUNCTION: Plays an essential role in type IV pili and type II
CC pseudopili formation by proteolytically removing the leader sequence
CC from substrate proteins and subsequently monomethylating the alpha-
CC amino group of the newly exposed N-terminal phenylalanine
CC (PubMed:8096341, PubMed:8340405, PubMed:23255525). Substrates include
CC proteins required for pilus biogenesis PilE, PilV, PilW, and PilX as
CC well as some components of the type II general secretory apparatus
CC GspG, GspH, GspI and GspJ (PubMed:8331069).
CC {ECO:0000269|PubMed:23255525, ECO:0000269|PubMed:8096341,
CC ECO:0000269|PubMed:8331069, ECO:0000269|PubMed:8340405}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Typically cleaves a -Gly-|-Phe- bond to release an N-terminal,
CC basic peptide of 5-8 residues from type IV prepilin, and then N-
CC methylates the new N-terminal amino group, the methyl donor being S-
CC adenosyl-L-methionine.; EC=3.4.23.43;
CC Evidence={ECO:0000269|PubMed:23255525, ECO:0000269|PubMed:8096341};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Note=Zinc is required for the N-terminal methylation of the mature
CC pilin, but not for signal peptide cleavage.
CC {ECO:0000269|PubMed:23255525};
CC -!- SUBCELLULAR LOCATION: Cell inner membrane {ECO:0000269|PubMed:8331069};
CC Multi-pass membrane protein.
CC -!- SIMILARITY: Belongs to the peptidase A24 family. {ECO:0000305}.
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DR EMBL; M32066; AAA25734.1; -; Genomic_DNA.
DR EMBL; M61096; AAA26023.1; -; Genomic_DNA.
DR EMBL; AE004091; AAG07916.1; -; Genomic_DNA.
DR PIR; A39131; A39131.
DR RefSeq; NP_253218.1; NC_002516.2.
DR RefSeq; WP_003112839.1; NZ_QZGE01000004.1.
DR AlphaFoldDB; P22610; -.
DR STRING; 287.DR97_1709; -.
DR MEROPS; A24.001; -.
DR TCDB; 3.A.15.2.1; the outer membrane protein secreting main terminal branch (mtb) family.
DR PaxDb; P22610; -.
DR EnsemblBacteria; AAG07916; AAG07916; PA4528.
DR GeneID; 877861; -.
DR KEGG; pae:PA4528; -.
DR PATRIC; fig|208964.12.peg.4739; -.
DR PseudoCAP; PA4528; -.
DR HOGENOM; CLU_057101_0_0_6; -.
DR InParanoid; P22610; -.
DR OMA; VFWLFKL; -.
DR PhylomeDB; P22610; -.
DR BioCyc; MetaCyc:MON-21661; -.
DR BioCyc; PAER208964:G1FZ6-4618-MON; -.
DR BRENDA; 3.4.23.43; 5087.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0005886; C:plasma membrane; IDA:PseudoCAP.
DR GO; GO:0015627; C:type II protein secretion system complex; IDA:PseudoCAP.
DR GO; GO:0004190; F:aspartic-type endopeptidase activity; IDA:PseudoCAP.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008170; F:N-methyltransferase activity; IDA:PseudoCAP.
DR GO; GO:0006480; P:N-terminal protein amino acid methylation; IDA:PseudoCAP.
DR GO; GO:0009297; P:pilus assembly; IDA:PseudoCAP.
DR GO; GO:0006465; P:signal peptide processing; IDA:PseudoCAP.
DR GO; GO:0043683; P:type IV pilus assembly; IMP:PseudoCAP.
DR InterPro; IPR010627; Pept_A24A_N.
DR InterPro; IPR014032; Peptidase_A24A_bac.
DR InterPro; IPR000045; Prepilin_IV_endopep_pep.
DR Pfam; PF06750; DiS_P_DiS; 1.
DR Pfam; PF01478; Peptidase_A24; 1.
DR PRINTS; PR00864; PREPILNPTASE.
PE 1: Evidence at protein level;
KW Cell inner membrane; Cell membrane; Hydrolase; Membrane; Metal-binding;
KW Methyltransferase; Multifunctional enzyme; Protease; Reference proteome;
KW S-adenosyl-L-methionine; Transferase; Transmembrane; Transmembrane helix;
KW Zinc.
FT CHAIN 1..290
FT /note="Prepilin leader peptidase/N-methyltransferase"
FT /id="PRO_0000192625"
FT TRANSMEM 13..33
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 128..148
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 158..178
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 183..203
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 228..248
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 261..276
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 72
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000305|PubMed:23255525"
FT BINDING 75
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000305|PubMed:23255525"
FT BINDING 97
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000305|PubMed:23255525"
FT BINDING 100
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000305|PubMed:23255525"
FT VARIANT 18
FT /note="T -> A (in strain: PAK)"
FT MUTAGEN 72
FT /note="C->G: About 80% loss of peptidase activity and
FT complete loss of methylase activity."
FT /evidence="ECO:0000269|PubMed:8340405"
FT MUTAGEN 75
FT /note="C->G: About 90% loss of peptidase activity and
FT complete loss of methylase activity."
FT /evidence="ECO:0000269|PubMed:8340405"
FT MUTAGEN 95
FT /note="G->D: Complete loss of methylase activity."
FT /evidence="ECO:0000269|PubMed:9668097"
FT MUTAGEN 96
FT /note="K->E: About 95% loss of methylase activity."
FT /evidence="ECO:0000269|PubMed:9668097"
FT MUTAGEN 97
FT /note="C->G: Complete loss of peptidase and methylase
FT activity."
FT /evidence="ECO:0000269|PubMed:8340405"
FT MUTAGEN 100
FT /note="C->G: About 95% loss of peptidase activity and
FT complete loss of methylase activity."
FT /evidence="ECO:0000269|PubMed:8340405"
SQ SEQUENCE 290 AA; 31870 MW; 57A8526EB18FB752 CRC64;
MPLLDYLASH PLAFVLCTIL LGLLVGSFLN VVVHRLPKMM ERNWKAEARE ALGLEPEPKQ
ATYNLVLPNS ACPRCGHEIR PWENIPLVSY LALGGKCSSC KAAIGKRYPL VELATALLSG
YVAWHFGFTW QAGAMLLLTW GLLAMSLIDA DHQLLPDVLV LPLLWLGLIA NHFGLFASLD
DALFGAVFGY LSLWSVFWLF KLVTGKEGMG YGDFKLLAML GAWGGWQILP LTILLSSLVG
AILGVIMLRL RNAESGTPIP FGPYLAIAGW IALLWGDQIT RTYLQFAGFK
