FEMX_WEIVI
ID FEMX_WEIVI Reviewed; 336 AA.
AC Q9EY50;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 81.
DE RecName: Full=UDP-N-acetylmuramoylpentapeptide-lysine N(6)-alanyltransferase;
DE EC=2.3.2.10 {ECO:0000269|PubMed:11083873, ECO:0000269|PubMed:4248527};
GN Name=femX {ECO:0000303|PubMed:11083873};
OS Weissella viridescens (Lactobacillus viridescens).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Weissella.
OX NCBI_TaxID=1629 {ECO:0000312|EMBL:AAG21689.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-24; 161-169 AND
RP 316-333, FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF GLN-29; TYR-73;
RP PRO-110; GLN-144; PHE-196; TYR-216; GLY-292; PHE-305; LYS-306 AND GLY-319.
RX PubMed=11083873; DOI=10.1074/jbc.m008591200;
RA Hegde S.S., Shrader T.E.;
RT "FemABX family members are novel nonribosomal peptidyltransferases and
RT important pathogen-specific drug targets.";
RL J. Biol. Chem. 276:6998-7003(2001).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=4248527; DOI=10.1016/s0021-9258(18)62979-5;
RA Plapp R., Strominger J.L.;
RT "Biosynthesis of the peptidoglycan of bacterial cell walls. 18.
RT Purification and properties of L-alanyl transfer ribonucleic acid-uridine
RT diphosphate-N-acetylmuramyl-pentapeptide transferase from Lactobacillus
RT viridescens.";
RL J. Biol. Chem. 245:3675-3682(1970).
RN [3]
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF ASP-109;
RP GLU-205; GLU-215; GLU-316 AND GLU-320.
RX PubMed=12679335; DOI=10.1074/jbc.m301565200;
RA Hegde S.S., Blanchard J.S.;
RT "Kinetic and mechanistic characterization of recombinant Lactobacillus
RT viridescens FemX (UDP-N-acetylmuramoyl pentapeptide-lysine N6-
RT alanyltransferase).";
RL J. Biol. Chem. 278:22861-22867(2003).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF 2-336 IN COMPLEX WITH SUBSTRATE.
RX PubMed=14962386; DOI=10.1016/j.str.2004.01.006;
RA Biarrotte-Sorin S., Maillard A.P., Delettre J., Sougakoff W., Arthur M.,
RA Mayer C.;
RT "Crystal structures of Weissella viridescens FemX and its complex with UDP-
RT MurNAc-pentapeptide: insights into FemABX family substrates recognition.";
RL Structure 12:257-267(2004).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF 2-336, FUNCTION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF LYS-37; ARG-212 AND
RP TYR-216.
RX PubMed=15901708; DOI=10.1128/jb.187.11.3833-3838.2005;
RA Maillard A.P., Biarrotte-Sorin S., Villet R., Mesnage S., Bouhss A.,
RA Sougakoff W., Mayer C., Arthur M.;
RT "Structure-based site-directed mutagenesis of the UDP-MurNAc-pentapeptide-
RT binding cavity of the FemX alanyl transferase from Weissella viridescens.";
RL J. Bacteriol. 187:3833-3838(2005).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS).
RA Delfosse V., Piton J., Villet R., Lecerf M., Arthur M., Mayer C.;
RT "Structure of Weissella viridescens FemX with the UDP-MurNAc-hexapeptide
RT product of the alanine transfer reaction.";
RL Submitted (MAR-2009) to the PDB data bank.
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.66 ANGSTROMS), FUNCTION, AND MUTAGENESIS OF
RP LYS-37; ASN-39; TRP-40; PHE-71; TYR-104; HIS-140; HIS-207; ILE-209;
RP THR-210; ARG-212; TYR-216; TYR-257; PHE-305 AND LYS-306.
RX PubMed=23744707; DOI=10.1002/anie.201301411;
RA Fonvielle M., Li de La Sierra-Gallay I., El-Sagheer A.H., Lecerf M.,
RA Patin D., Mellal D., Mayer C., Blanot D., Gale N., Brown T.,
RA van Tilbeurgh H., Etheve-Quelquejeu M., Arthur M.;
RT "The structure of FemX(Wv) in complex with a peptidyl-RNA conjugate:
RT mechanism of aminoacyl transfer from Ala-tRNA(Ala) to peptidoglycan
RT precursors.";
RL Angew. Chem. Int. Ed. 52:7278-7281(2013).
CC -!- FUNCTION: Involved in the synthesis of the bacterial cell wall.
CC Catalyzes the addition of alanine into the interchain peptide bridge of
CC peptidoglycan precursor using aminoacyl-tRNA(Ala) as amino acid donor.
CC This alanine is added to the epsilon-amino group of the L-lysine of the
CC peptidoglycan UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-L-
CC lysyl-D-alanyl-D-alanine, in a ribosome-independent mechanism
CC (PubMed:11083873, PubMed:4248527, PubMed:12679335, PubMed:15901708,
CC PubMed:23744707). Specific for UDP-N-acetyl-muramoyl-pentapeptide. Has
CC no activity toward UDP-N-acetyl-muramoyl-tetrapeptide or UDP-N-acetyl-
CC muramoyl-tripeptide (PubMed:15901708). Also acts on L-seryl-tRNA(Ser)
CC (PubMed:4248527). {ECO:0000269|PubMed:11083873,
CC ECO:0000269|PubMed:12679335, ECO:0000269|PubMed:15901708,
CC ECO:0000269|PubMed:23744707, ECO:0000269|PubMed:4248527}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-alanyl-tRNA(Ala) + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-
CC gamma-D-glutamyl-L-lysyl-D-alanyl-D-alanine = H(+) + tRNA(Ala) + UDP-
CC N-acetyl-alpha-D-muramoyl-L-alanyl-gamma-D-glutamyl-N(6)-(L-alanyl)-
CC L-lysyl-D-alanyl-D-alanine; Xref=Rhea:RHEA:12432, Rhea:RHEA-
CC COMP:9657, Rhea:RHEA-COMP:9923, ChEBI:CHEBI:15378, ChEBI:CHEBI:70758,
CC ChEBI:CHEBI:71358, ChEBI:CHEBI:78442, ChEBI:CHEBI:78497; EC=2.3.2.10;
CC Evidence={ECO:0000269|PubMed:11083873, ECO:0000269|PubMed:4248527};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=15 uM for tRNA(Ala) {ECO:0000269|PubMed:12679335};
CC KM=1.7 uM for tRNA(Ala) {ECO:0000269|PubMed:15901708};
CC KM=42 uM for UDP-N-acetyl-muramoyl-pentapeptide
CC {ECO:0000269|PubMed:12679335};
CC KM=79 uM for UDP-N-acetyl-muramoyl-pentapeptide
CC {ECO:0000269|PubMed:15901708};
CC pH dependence:
CC Optimum pH is 7.0-8.0. {ECO:0000269|PubMed:12679335};
CC -!- SIMILARITY: Belongs to the FemABX family. {ECO:0000305}.
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DR EMBL; AY008262; AAG21689.1; -; Genomic_DNA.
DR RefSeq; WP_057745451.1; NZ_JQBM01000002.1.
DR PDB; 1NE9; X-ray; 1.70 A; A=2-336.
DR PDB; 1P4N; X-ray; 1.90 A; A=2-336.
DR PDB; 1XE4; X-ray; 1.95 A; A=2-336.
DR PDB; 1XF8; X-ray; 1.90 A; A=2-336.
DR PDB; 1XIX; X-ray; 2.00 A; A=2-336.
DR PDB; 3GKR; X-ray; 1.60 A; A=1-336.
DR PDB; 4II9; X-ray; 1.66 A; A=1-336.
DR PDBsum; 1NE9; -.
DR PDBsum; 1P4N; -.
DR PDBsum; 1XE4; -.
DR PDBsum; 1XF8; -.
DR PDBsum; 1XIX; -.
DR PDBsum; 3GKR; -.
DR PDBsum; 4II9; -.
DR AlphaFoldDB; Q9EY50; -.
DR SMR; Q9EY50; -.
DR STRING; 1629.IV50_GL000816; -.
DR KEGG; ag:AAG21689; -.
DR OMA; FGLIQFK; -.
DR OrthoDB; 891566at2; -.
DR BioCyc; MetaCyc:MON-15493; -.
DR BRENDA; 2.3.2.10; 2901.
DR BRENDA; 2.3.2.16; 2901.
DR SABIO-RK; Q9EY50; -.
DR EvolutionaryTrace; Q9EY50; -.
DR GO; GO:0047206; F:UDP-N-acetylmuramoylpentapeptide-lysine N6-alanyltransferase activity; IDA:UniProtKB.
DR GO; GO:0071555; P:cell wall organization; IEA:UniProtKB-KW.
DR GO; GO:0009252; P:peptidoglycan biosynthetic process; IDA:UniProtKB.
DR GO; GO:0008360; P:regulation of cell shape; IEA:UniProtKB-KW.
DR InterPro; IPR016181; Acyl_CoA_acyltransferase.
DR InterPro; IPR003447; FEMABX.
DR Pfam; PF02388; FemAB; 2.
DR SUPFAM; SSF55729; SSF55729; 2.
DR PROSITE; PS51191; FEMABX; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Cell shape;
KW Cell wall biogenesis/degradation; Direct protein sequencing;
KW Peptidoglycan synthesis; Transferase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:11083873"
FT CHAIN 2..336
FT /note="UDP-N-acetylmuramoylpentapeptide-lysine N(6)-
FT alanyltransferase"
FT /id="PRO_0000430779"
FT BINDING 37..40
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14962386"
FT BINDING 104
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14962386"
FT BINDING 212
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14962386"
FT BINDING 216
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14962386"
FT BINDING 257
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:14962386"
FT SITE 109
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000303|PubMed:12679335"
FT SITE 320
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000303|PubMed:12679335"
FT MUTAGEN 29
FT /note="Q->E: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 29
FT /note="Q->T: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 37
FT /note="K->M: Loss of activity."
FT /evidence="ECO:0000269|PubMed:15901708,
FT ECO:0000269|PubMed:23744707"
FT MUTAGEN 37
FT /note="K->R: Reduces activity 150-fold."
FT /evidence="ECO:0000269|PubMed:15901708"
FT MUTAGEN 37
FT /note="K->R: Reduces activity 200-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 39
FT /note="N->L: No effect on activity."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 40
FT /note="W->F: Reduces activity 2-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 71
FT /note="F->L: Reduces activity 2-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 73
FT /note="Y->F: Reduces activity 2.5-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 73
FT /note="Y->L: Reduces activity 2.5-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 104
FT /note="Y->F: No effect on activity."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 109
FT /note="D->N: Reduces activity 200-fold."
FT /evidence="ECO:0000269|PubMed:12679335"
FT MUTAGEN 110
FT /note="P->F: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 110
FT /note="P->H: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 140
FT /note="H->A: Reduces activity 5-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 144
FT /note="Q->E: Reduces activity 9-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 144
FT /note="Q->T: Slightly reduces activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 196
FT /note="F->L: Reduces activity 4-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 196
FT /note="F->Y: Slightly reduces activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 205
FT /note="E->Q: No effect on activity."
FT /evidence="ECO:0000269|PubMed:12679335"
FT MUTAGEN 207
FT /note="H->A: Reduces activity 11-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 209
FT /note="I->A: Reduces activity 50-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 210
FT /note="T->A: Reduces activity 2-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 212
FT /note="R->K: Loss of activity."
FT /evidence="ECO:0000269|PubMed:15901708"
FT MUTAGEN 212
FT /note="R->M: Loss of activity."
FT /evidence="ECO:0000269|PubMed:15901708,
FT ECO:0000269|PubMed:23744707"
FT MUTAGEN 215
FT /note="E->Q: No effect on activity."
FT /evidence="ECO:0000269|PubMed:12679335"
FT MUTAGEN 216
FT /note="Y->F: Reduced activity."
FT /evidence="ECO:0000269|PubMed:11083873,
FT ECO:0000269|PubMed:15901708"
FT MUTAGEN 216
FT /note="Y->F: Reduces activity 25-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 216
FT /note="Y->L: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 257
FT /note="Y->F: Reduces activity 2-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 292
FT /note="G->V: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 305
FT /note="F->A: Reduces activity 17-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 305
FT /note="F->L: Reduces activity 33-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 305
FT /note="F->L: Reduces activity 6-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 305
FT /note="F->Y: No effect on activity."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 306
FT /note="K->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 306
FT /note="K->A: Reduces activity 12-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 306
FT /note="K->C: Reduces activity 2.5-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 306
FT /note="K->M: Reduces activity 200-fold."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 306
FT /note="K->N: Reduces activity 11-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 306
FT /note="K->R: Loss of activity."
FT /evidence="ECO:0000269|PubMed:23744707"
FT MUTAGEN 316
FT /note="E->Q: Reduces activity 3-fold."
FT /evidence="ECO:0000269|PubMed:12679335"
FT MUTAGEN 319
FT /note="G->S: Reduces activity 8-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 319
FT /note="G->V: Reduces activity 20-fold."
FT /evidence="ECO:0000269|PubMed:11083873"
FT MUTAGEN 320
FT /note="E->Q: Reduces activity 25-fold."
FT /evidence="ECO:0000269|PubMed:12679335"
FT HELIX 9..21
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 27..29
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 31..36
FT /evidence="ECO:0007829|PDB:3GKR"
FT TURN 37..39
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 40..48
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 54..63
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 66..74
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 85..99
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 103..108
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 116..124
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 128..131
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 132..134
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 136..138
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 139..141
FT /evidence="ECO:0007829|PDB:4II9"
FT STRAND 142..144
FT /evidence="ECO:0007829|PDB:4II9"
FT STRAND 146..152
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 153..155
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 162..165
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 168..179
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 182..187
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 190..207
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 214..223
FT /evidence="ECO:0007829|PDB:3GKR"
FT TURN 226..228
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 229..236
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 239..249
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 252..260
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 268..282
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 286..292
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 301..309
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 315..317
FT /evidence="ECO:0007829|PDB:3GKR"
FT STRAND 320..325
FT /evidence="ECO:0007829|PDB:3GKR"
FT HELIX 327..333
FT /evidence="ECO:0007829|PDB:3GKR"
SQ SEQUENCE 336 AA; 38284 MW; AFB94A2718148C3C CRC64;
MPVLNLNDPQ AVERYEEFMR QSPYGQVTQD LGWAKVKNNW EPVDVYLEDD QGAIIAAMSM
LLGDTPTDKK FAYASKGPVM DVTDVDLLDR LVDEAVKALD GRAYVLRFDP EVAYSDEFNT
TLQDHGYVTR NRNVADAGMH ATIQPRLNMV LDLTKFPDAK TTLDLYPSKT KSKIKRPFRD
GVEVHSGNSA TELDEFFKTY TTMAERHGIT HRPIEYFQRM QAAFDADTMR IFVAEREGKL
LSTGIALKYG RKIWYMYAGS MDGNTYYAPY AVQSEMIQWA LDTNTDLYDL GGIESESTDD
SLYVFKHVFV KDAPREYIGE IDKVLDPEVY AELVKD
