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PDBsum entry 1cl1

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Methionine biosynthesis PDB id
1cl1
Jmol
Contents
Protein chains
391 a.a. *
Ligands
BCT ×2
Waters ×1875
* Residue conservation analysis
PDB id:
1cl1
Name: Methionine biosynthesis
Title: Cystathionine beta-lyase (cbl) from escherichia coli
Structure: Cystathionine beta-lyase. Chain: a, b. Synonym: beta cystathionase. Engineered: yes. Other_details: plp bound as cofactor to lys 210
Source: Escherichia coli. Organism_taxid: 83333. Strain: k12. Gene: metc. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Tetramer (from PQS)
Resolution:
1.83Å     R-factor:   0.151     R-free:   0.221
Authors: T.Clausen,R.Huber,A.Messerschmidt
Key ref:
T.Clausen et al. (1996). Crystal structure of the pyridoxal-5'-phosphate dependent cystathionine beta-lyase from Escherichia coli at 1.83 A. J Mol Biol, 262, 202-224. PubMed id: 8831789 DOI: 10.1006/jmbi.1996.0508
Date:
02-Sep-97     Release date:   09-Sep-98    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P06721  (METC_ECOLI) -  Cystathionine beta-lyase MetC
Seq:
Struc:
395 a.a.
391 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.4.4.1.8  - Cystathionine beta-lyase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: L-cystathionine + H2O = L-homocysteine + NH3 + pyruvate
L-cystathionine
+ H(2)O
= L-homocysteine
+ NH(3)
+
pyruvate
Bound ligand (Het Group name = BCT)
matches with 42.86% similarity
      Cofactor: Pyridoxal 5'-phosphate
Pyridoxal 5'-phosphate
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   1 term 
  Biological process     cellular amino acid metabolic process   3 terms 
  Biochemical function     catalytic activity     5 terms  

 

 
    reference    
 
 
DOI no: 10.1006/jmbi.1996.0508 J Mol Biol 262:202-224 (1996)
PubMed id: 8831789  
 
 
Crystal structure of the pyridoxal-5'-phosphate dependent cystathionine beta-lyase from Escherichia coli at 1.83 A.
T.Clausen, R.Huber, B.Laber, H.D.Pohlenz, A.Messerschmidt.
 
  ABSTRACT  
 
Cystathionine beta-lyase (CBL) is a member of the gamma-family of PLP-dependent enzymes, that cleaves C beta-S bonds of a broad variety of substrates. The crystal structure of CBL from E. coli has been solved using MIR phases in combination with density modification. The structure has been refined to an R-factor of 15.2% at 1.83 A resolution using synchroton radiation diffraction data. The asymmetric unit of the crystal cell (space group C222(1)) contains two monomers related by 2-fold symmetry. A homotetramer with 222 symmetry is built up by crystallographic and non-crystallographic symmetry. Each monomer of CBL can be described in terms of three spatially and functionally different domains. The N-terminal domain (residues 1 to 60) consists of three alpha-helices and one beta-strand. It contributes to tetramer formation and is part of the active site of the adjacent subunit. The second domain (residues 61 to 256) harbors PLP and has an alpha/beta-structure with a seven-stranded beta-sheet as the central part. The remaining C-terminal domain (residues 257 to 395), connected by a long alpha-helix to the PLP-binding domain, consists of four helices packed on the solvent-accessible side of an antiparallel four-stranded beta-sheet. The fold of the C-terminal and the PLP-binding domain and the location of the active site are similar to aminotransferases. Most of the residues in the active site are strongly conserved among the enzymes of the transsulfuration pathway. Additionally, CBL is homologous to the mal gamma gene product indicating an evolutionary relationship between alpha and gamma-family of PLP-dependent enzymes. The structure of the beta, beta, beta-trifluoroalanine inactivated CBL has been refined at 2.3 A resolution to an R-factor of 16.2%. It suggests that Lys210, the PLP-binding residue, mediates the proton transfer between C alpha and S gamma.
 
  Selected figure(s)  
 
Figure 11.
Figure 11. Proposed reaction mechanism (A)--(F) for inactivation of CBL by F3Ala. Lys210 is the nucleophile at the active site that reacts via Michael addition with the so-called activated Michael acceptor (C). Subsequent elimination of HF from C b results in the inactivation end product (F) which was observed in the crystal structure of the CBL--F3Ala complex. Abbreviation used: Pyr-C = NH + , enzyme-bound PLP.
Figure 13.
Figure 13. Proposed reaction mechanism for CBL. The colors used are: apoprotein, red; cofactor, black; substrate, blue. (a) Residues involved in substrate binding. (b) Residues essential for catalysis. (c) Mechanism of C b -S bond cleavage (Pyr-C = NH + , PLP derivative).
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (1996, 262, 202-224) copyright 1996.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

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High-resolution structure of methionine gamma-lyase from Citrobacter freundii.
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PDB code: 2rfv
18603802 D.Kudou, S.Misaki, M.Yamashita, T.Tamura, N.Esaki, and K.Inagaki (2008).
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PDB code: 2vgz
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PDB codes: 3bc8 3bca 3bcb
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PDB code: 2g6w
16820483 M.C.Martínez-Cuesta, C.Peláez, J.Eagles, M.J.Gasson, T.Requena, and S.B.Hanniffy (2006).
YtjE from Lactococcus lactis IL1403 Is a C-S lyase with alpha, gamma-elimination activity toward methionine.
  Appl Environ Microbiol, 72, 4878-4884.  
  16511092 D.V.Mamaeva, E.A.Morozova, A.D.Nikulin, S.V.Revtovich, S.V.Nikonov, M.B.Garber, and T.V.Demidkina (2005).
Structure of Citrobacter freundii L-methionine gamma-lyase.
  Acta Crystallogr Sect F Struct Biol Cryst Commun, 61, 546-549.
PDB code: 1y4i
16307301 M.Wirtz, and M.Droux (2005).
Synthesis of the sulfur amino acids: cysteine and methionine.
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16000837 N.Awano, M.Wada, H.Mori, S.Nakamori, and H.Takagi (2005).
Identification and functional analysis of Escherichia coli cysteine desulfhydrases.
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15498941 A.Paiardini, F.Bossa, and S.Pascarella (2004).
Evolutionarily conserved regions and hydrophobic contacts at the superfamily level: The case of the fold-type I, pyridoxal-5'-phosphate-dependent enzymes.
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15044494 C.R.Sweet, A.A.Ribeiro, and C.R.Raetz (2004).
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15155634 L.J.Ejim, V.M.D'Costa, N.H.Elowe, J.C.Loredo-Osti, D.Malo, and G.D.Wright (2004).
Cystathionine beta-lyase is important for virulence of Salmonella enterica serovar Typhimurium.
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12715888 A.Messerschmidt, M.Worbs, C.Steegborn, M.C.Wahl, R.Huber, B.Laber, and T.Clausen (2003).
Determinants of enzymatic specificity in the Cys-Met-metabolism PLP-dependent enzymes family: crystal structure of cystathionine gamma-lyase from yeast and intrafamiliar structure comparison.
  Biol Chem, 384, 373-386.
PDB code: 1n8p
12882962 T.Ose, A.Fujino, M.Yao, N.Watanabe, M.Honma, and I.Tanaka (2003).
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PDB codes: 1j0c 1j0d 1j0e
12191993 J.Zhang, and G.C.Ferreira (2002).
Transient state kinetic investigation of 5-aminolevulinate synthase reaction mechanism.
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11756443 M.Garrido-Franco, S.Ehlert, A.Messerschmidt, S.Marinkovic', R.Huber, B.Laber, G.P.Bourenkov, and T.Clausen (2002).
Structure and function of threonine synthase from yeast.
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11933244 K.Soda, T.Yoshimura, and N.Esaki (2001).
Stereospecificity for the hydrogen transfer of pyridoxal enzyme reactions.
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The manifold of vitamin B6 dependent enzymes.
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10880431 H.I.Krupka, R.Huber, S.C.Holt, and T.Clausen (2000).
Crystal structure of cystalysin from Treponema denticola: a pyridoxal 5'-phosphate-dependent protein acting as a haemolytic enzyme.
  EMBO J, 19, 3168-3178.
PDB codes: 1c7n 1c7o
11193400 H.Inoue, K.Inagaki, N.Adachi, T.Tamura, N.Esaki, K.Soda, and H.Tanaka (2000).
Role of tyrosine 114 of L-methionine gamma-lyase from Pseudomonas putida.
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10618201 M.Fernández, W.van Doesburg, G.A.Rutten, J.D.Marugg, A.C.Alting, R.van Kranenburg, and O.P.Kuipers (2000).
Molecular and functional analyses of the metC gene of Lactococcus lactis, encoding cystathionine beta-lyase.
  Appl Environ Microbiol, 66, 42-48.  
  10620674 N.Dobric, G.K.Limsowtin, A.J.Hillier, N.P.Dudman, and B.E.Davidson (2000).
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10698925 T.Clausen, A.Schlegel, R.Peist, E.Schneider, C.Steegborn, Y.S.Chang, A.Haase, G.P.Bourenkov, H.D.Bartunik, and W.Boos (2000).
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  EMBO J, 19, 831-842.
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10684605 T.Fujii, M.Maeda, H.Mihara, T.Kurihara, N.Esaki, and Y.Hata (2000).
Structure of a NifS homologue: X-ray structure analysis of CsdB, an Escherichia coli counterpart of mammalian selenocysteine lyase.
  Biochemistry, 39, 1263-1273.
PDB code: 1c0n
11092940 V.Sridhar, M.Xu, Q.Han, X.Sun, Y.Tan, R.M.Hoffman, and G.S.Prasad (2000).
Crystallization and preliminary crystallographic characterization of recombinant L-methionine-alpha-deamino-gamma-mercaptomethane lyase (methioninase).
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10378276 A.D.Kern, M.A.Oliveira, P.Coffino, and M.L.Hackert (1999).
Structure of mammalian ornithine decarboxylase at 1.6 A resolution: stereochemical implications of PLP-dependent amino acid decarboxylases.
  Structure, 7, 567-581.
PDB code: 7odc
10584065 A.Poupon, F.Jebai, G.Labesse, F.Gros, J.Thibault, J.P.Mornon, and M.Krieger (1999).
Structure modelling and site-directed mutagenesis of the rat aromatic L-amino acid pyridoxal 5'-phosphate-dependent decarboxylase: a functional study.
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10212249 C.Steegborn, T.Clausen, P.Sondermann, U.Jacob, M.Worbs, S.Marinkovic, R.Huber, and M.C.Wahl (1999).
Kinetics and inhibition of recombinant human cystathionine gamma-lyase. Toward the rational control of transsulfuration.
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Rat liver serine dehydratase. Bacterial expression and two folding domains as revealed by limited proteolysis.
  J Biol Chem, 274, 12855-12860.  
10509022 J.Brzywczy, and A.Paszewski (1999).
Cloning and characterization of the Kluyveromyces lactis homocysteine synthase gene.
  Yeast, 15, 1403-1409.  
10382666 P.K.Mehta, P.Argos, A.D.Barbour, and P.Christen (1999).
Recognizing very distant sequence relationships among proteins by family profile analysis.
  Proteins, 35, 387-400.  
10595588 T.Clausen, M.C.Wahl, A.Messerschmidt, R.Huber, J.C.Fuhrmann, B.Laber, W.Streber, and C.Steegborn (1999).
Cloning, purification and characterisation of cystathionine gamma-synthase from Nicotiana tabacum.
  Biol Chem, 380, 1237-1242.  
9488680 A.E.McKie, T.Edlind, J.Walker, J.C.Mottram, and G.H.Coombs (1998).
The primitive protozoon Trichomonas vaginalis contains two methionine gamma-lyase genes that encode members of the gamma-family of pyridoxal 5'-phosphate-dependent enzymes.
  J Biol Chem, 273, 5549-5556.  
9914259 J.N.Jansonius (1998).
Structure, evolution and action of vitamin B6-dependent enzymes.
  Curr Opin Struct Biol, 8, 759-769.  
9772188 K.H.Jhee, P.McPhie, H.S.Ro, and E.W.Miles (1998).
Tryptophan synthase mutations that alter cofactor chemistry lead to mechanism-based inactivation.
  Biochemistry, 37, 14591-14604.  
  9605314 K.Qu, D.L.Martin, and C.E.Lawrence (1998).
Motifs and structural fold of the cofactor binding site of human glutamate decarboxylase.
  Protein Sci, 7, 1092-1105.  
9753690 S.B.Renwick, K.Snell, and U.Baumann (1998).
The crystal structure of human cytosolic serine hydroxymethyltransferase: a target for cancer chemotherapy.
  Structure, 6, 1105-1116.
PDB code: 1bj4
  10082378 S.Ishii, H.Hayashi, A.Okamoto, and H.Kagamiyama (1998).
Aromatic L-amino acid decarboxylase: conformational change in the flexible region around Arg334 is required during the transaldimination process.
  Protein Sci, 7, 1802-1810.  
9843488 T.Clausen, R.Huber, L.Prade, M.C.Wahl, and A.Messerschmidt (1998).
Crystal structure of Escherichia coli cystathionine gamma-synthase at 1.5 A resolution.
  EMBO J, 17, 6827-6838.
PDB code: 1cs1
  9529892 W.Boos, and H.Shuman (1998).
Maltose/maltodextrin system of Escherichia coli: transport, metabolism, and regulation.
  Microbiol Mol Biol Rev, 62, 204-229.  
  9165087 L.Prade, P.Hof, and B.Bieseler (1997).
Dimer interface of glutathione S-transferase from Arabidopsis thaliana: influence of the G-site architecture on the dimer interface and implications for classification.
  Biol Chem, 378, 317-320.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.