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Q05514

- MAAL_CLOTT

UniProt

Q05514 - MAAL_CLOTT

Protein

Methylaspartate ammonia-lyase

Gene
N/A
Organism
Clostridium tetanomorphum
Status
Reviewed - Annotation score: 5 out of 5- Experimental evidence at protein leveli
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    • History
      Entry version 75 (01 Oct 2014)
      Sequence version 1 (01 Feb 1995)
      Previous versions | rss
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    Functioni

    Involved in the methylaspartate cycle. Catalyzes the formation of the alpha,beta-unsaturated bond by the reversible anti elimination of ammonia from L-threo-beta-methylaspartate (L-threo-(2S,3S)-3-methylaspartate) to give mesaconate. It can also use L-erythro-beta-methylaspartate (L-erythro-(2S,3R)-3-methylaspartate), L-aspartate, fumarate and ethylfumarate as substrates.4 Publications

    Catalytic activityi

    L-threo-3-methylaspartate = mesaconate + NH3.4 Publications

    Cofactori

    Magnesium.3 Publications

    Enzyme regulationi

    Inhibited by calcium ions.1 Publication

    Kineticsi

    Kcat is 61 sec(-1) for amination of mesaconate (with 20 mM of MgCl2 at pH 9 and at 30 degrees Celsius). Kcat is 89 sec(-1) for deamination of L-threo-beta-methylaspartate (with 20 mM of MgCl2 at pH 9 and at 30 degrees Celsius).

    1. KM=0.65 mM for L-threo-beta-methylaspartate (with 4 mM of KCl at pH 9.76 and at 25 degrees Celsius)3 Publications
    2. KM=0.67 mM for L-threo-beta-methylaspartate (with 50 mM of KCl at pH 9 and at 30 degrees Celsius)3 Publications
    3. KM=0.7 mM for mesaconate (with 20 mM of MgCl2 at pH 9 and at 30 degrees Celsius)3 Publications
    4. KM=1 mM for L-threo-beta-methylaspartate (with 20 mM of MgCl2 at pH 9 and at 30 degrees Celsius)3 Publications
    5. KM=2.3 mM for L-aspartate (with 4 mM of KCl at pH 9.76 and at 25 degrees Celsius)3 Publications
    6. KM=2.8 mM for L-threo-beta-methylaspartate (with 0.3 mM of KCl at pH 9 and at 30 degrees Celsius)3 Publications

    Vmax=2089 µmol/min/mg enzyme with L-threo-beta-methylaspartate as substrate (with 50 mM of KCl at pH 9 and at 30 degrees Celsius)3 Publications

    Vmax=309 µmol/min/mg enzyme with L-threo-beta-methylaspartate as substrate (with 0.3 mM of KCl at pH 9 and at 30 degrees Celsius)3 Publications

    Vmax=266 µmol/min/mg enzyme with L-threo-beta-methylaspartate as substrate (with 4 mM of KCl at pH 9.76 and at 25 degrees Celsius)3 Publications

    Vmax=2.5 µmol/min/mg enzyme with L-erythro-beta-methylaspartate as substrate (with 4 mM of KCl at pH 9.76 and at 25 degrees Celsius)3 Publications

    Vmax=2.4 µmol/min/mg enzyme with L-aspartate as substrate (with 4 mM of KCl at pH 9.76 and at 25 degrees Celsius)3 Publications

    pH dependencei

    Optimum pH is 9.7.3 Publications

    Temperature dependencei

    Optimum temperature is 55 degrees Celsius. It retains only half of its original activity after a 30 minutes incubation period at 50 degrees Celsius.3 Publications

    Pathwayi

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei172 – 1721L-threo-beta-methylaspartateBy similarity
    Sitei194 – 1941Transition state stabilizer
    Metal bindingi238 – 2381Magnesium2 Publications
    Metal bindingi273 – 2731Magnesium2 Publications
    Metal bindingi307 – 3071Magnesium2 Publications
    Binding sitei329 – 3291L-threo-beta-methylaspartate
    Active sitei331 – 3311Proton acceptor2 Publications
    Binding sitei361 – 3611L-threo-beta-methylaspartate

    GO - Molecular functioni

    1. cobalamin binding Source: UniProtKB-KW
    2. metal ion binding Source: UniProtKB-KW
    3. methylaspartate ammonia-lyase activity Source: UniProtKB-EC

    GO - Biological processi

    1. glutamate catabolic process via L-citramalate Source: UniProtKB-UniPathway

    Keywords - Molecular functioni

    Lyase

    Keywords - Ligandi

    Cobalamin, Cobalt, Magnesium, Metal-binding

    Enzyme and pathway databases

    BioCyciMetaCyc:MONOMER-1103.
    UniPathwayiUPA00561; UER00618.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Methylaspartate ammonia-lyase (EC:4.3.1.2)
    Short name:
    MAL
    Alternative name(s):
    3-methylaspartase ammonia-lyase
    Beta-methylaspartase
    OrganismiClostridium tetanomorphum
    Taxonomic identifieri1553 [NCBI]
    Taxonomic lineageiBacteriaFirmicutesClostridiaClostridialesClostridiaceaeClostridium

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi73 – 731Q → A: It has very broad nucleophile scope and excellent regio- and diastereoselectivity in the amination reaction. This mutation strongly moves the specificity of MAL away from ammonia and towards methylamine. It is highly enantioselective. 1 Publication
    Mutagenesisi194 – 1941H → A: Strong (160-fold) decrease of the catalytic efficiency for deamination and slight (1.8-fold) decrease of affinity binding for L-threo-beta-methylaspartate. 7-fold decrease of the catalytic efficiency for amination and 20-fold decrease of affinity binding for mesaconate. It does not show any major conformational changes. 1 Publication
    Mutagenesisi194 – 1941H → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi329 – 3291Q → A: Very strong decrease of the catalytic efficiency for deamination, whereas the affinity binding for L-threo-beta-methylaspartate is not affected. Strong (240-fold) decrease of the catalytic efficiency for amination and slight (2.4-fold) decrease of affinity binding for mesaconate. It does not show any major conformational changes. 1 Publication
    Mutagenesisi329 – 3291Q → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi331 – 3311K → A: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi331 – 3311K → G: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi331 – 3311K → H: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi331 – 3311K → Q: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi331 – 3311K → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication
    Mutagenesisi384 – 3841L → A: It has very broad electrophile scope and excellent regio- and enantioselectivity in the amination reaction. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 413413Methylaspartate ammonia-lyasePRO_0000084547Add
    BLAST

    Interactioni

    Subunit structurei

    Homodimer.4 Publications

    Structurei

    Secondary structure

    1
    413
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi2 – 1110
    Beta strandi14 – 185
    Helixi20 – 245
    Beta strandi28 – 303
    Beta strandi33 – 364
    Beta strandi44 – 496
    Beta strandi52 – 598
    Beta strandi64 – 696
    Turni73 – 764
    Helixi86 – 9611
    Helixi98 – 1014
    Helixi109 – 11810
    Helixi128 – 14619
    Helixi150 – 1589
    Helixi179 – 1868
    Beta strandi190 – 1945
    Helixi200 – 2045
    Helixi209 – 22517
    Beta strandi234 – 2385
    Helixi242 – 2465
    Turni247 – 2493
    Helixi251 – 26515
    Beta strandi270 – 2734
    Helixi281 – 29818
    Beta strandi302 – 3065
    Helixi313 – 3219
    Beta strandi325 – 3306
    Helixi333 – 3353
    Helixi339 – 35012
    Beta strandi354 – 3574
    Helixi365 – 37814
    Beta strandi381 – 3844
    Beta strandi387 – 3915
    Helixi392 – 41019

    3D structure databases

    Select the link destinations:
    PDBe
    RCSB PDB
    PDBj
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1KCZX-ray1.90A/B1-413[»]
    1KD0X-ray1.90A/B1-413[»]
    3ZVHX-ray1.99A/B1-413[»]
    3ZVIX-ray1.90A/B1-413[»]
    ProteinModelPortaliQ05514.
    SMRiQ05514. Positions 1-413.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiQ05514.

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni360 – 3612L-threo-beta-methylaspartate bindingBy similarity

    Sequence similaritiesi

    Family and domain databases

    Gene3Di3.20.20.120. 1 hit.
    3.30.390.10. 1 hit.
    InterProiIPR029065. Enolase_C-like.
    IPR029017. Enolase_N_like.
    IPR006395. Me_Asp_am_lyase.
    IPR022662. MeAsp_NH4-lyase_C.
    IPR022665. MeAsp_NH4-lyase_N.
    IPR001354. MR_MLE.
    [Graphical view]
    PANTHERiPTHR13794. PTHR13794. 1 hit.
    PfamiPF07476. MAAL_C. 1 hit.
    PF05034. MAAL_N. 1 hit.
    [Graphical view]
    PIRSFiPIRSF017107. MAL. 1 hit.
    SUPFAMiSSF51604. SSF51604. 1 hit.
    SSF54826. SSF54826. 1 hit.
    TIGRFAMsiTIGR01502. B_methylAsp_ase. 1 hit.

    Sequencei

    Sequence statusi: Complete.

    Q05514-1 [UniParc]FASTAAdd to Basket

    « Hide

    MKIVDVLCTP GLTGFYFDDQ RAIKKGAGHD GFTYTGSTVT EGFTQVRQKG    50
    ESISVLLVLE DGQVAHGDCA AVQYSGAGGR DPLFLAKDFI PVIEKEIAPK 100
    LIGREITNFK PMAEEFDKMT VNGNRLHTAI RYGITQAILD AVAKTRKVTM 150
    AEVIRDEYNP GAEINAVPVF AQSGDDRYDN VDKMIIKEAD VLPHALINNV 200
    EEKLGLKGEK LLEYVKWLRD RIIKLRVRED YAPIFHIDVY GTIGAAFDVD 250
    IKAMADYIQT LAEAAKPFHL RIEGPMDVED RQKQMEAMRD LRAELDGRGV 300
    DAELVADEWC NTVEDVKFFT DNKAGHMVQI KTPDLGGVNN IADAIMYCKA 350
    NGMGAYCGGT CNETNRSAEV TTNIGMACGA RQVLAKPGMG VDEGMMIVKN 400
    EMNRVLALVG RRK 413
    Length:413
    Mass (Da):45,534
    Last modified:February 1, 1995 - v1
    Checksum:i4451923DB035EF13
    GO

    Sequence databases

    Select the link destinations:
    EMBL
    GenBank
    DDBJ
    Links Updated
    S48141 Genomic DNA. Translation: AAB24070.1.
    X70499 Genomic DNA. Translation: CAA49911.1.
    X70695 Genomic DNA. Translation: CAA50027.1.
    PIRiB44285.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBL
    GenBank
    DDBJ
    Links Updated
    S48141 Genomic DNA. Translation: AAB24070.1 .
    X70499 Genomic DNA. Translation: CAA49911.1 .
    X70695 Genomic DNA. Translation: CAA50027.1 .
    PIRi B44285.

    3D structure databases

    Select the link destinations:
    PDBe
    RCSB PDB
    PDBj
    Links Updated
    Entry Method Resolution (Å) Chain Positions PDBsum
    1KCZ X-ray 1.90 A/B 1-413 [» ]
    1KD0 X-ray 1.90 A/B 1-413 [» ]
    3ZVH X-ray 1.99 A/B 1-413 [» ]
    3ZVI X-ray 1.90 A/B 1-413 [» ]
    ProteinModelPortali Q05514.
    SMRi Q05514. Positions 1-413.
    ModBasei Search...
    MobiDBi Search...

    Protocols and materials databases

    Structural Biology Knowledgebase Search...

    Enzyme and pathway databases

    UniPathwayi UPA00561 ; UER00618 .
    BioCyci MetaCyc:MONOMER-1103.

    Miscellaneous databases

    EvolutionaryTracei Q05514.

    Family and domain databases

    Gene3Di 3.20.20.120. 1 hit.
    3.30.390.10. 1 hit.
    InterProi IPR029065. Enolase_C-like.
    IPR029017. Enolase_N_like.
    IPR006395. Me_Asp_am_lyase.
    IPR022662. MeAsp_NH4-lyase_C.
    IPR022665. MeAsp_NH4-lyase_N.
    IPR001354. MR_MLE.
    [Graphical view ]
    PANTHERi PTHR13794. PTHR13794. 1 hit.
    Pfami PF07476. MAAL_C. 1 hit.
    PF05034. MAAL_N. 1 hit.
    [Graphical view ]
    PIRSFi PIRSF017107. MAL. 1 hit.
    SUPFAMi SSF51604. SSF51604. 1 hit.
    SSF54826. SSF54826. 1 hit.
    TIGRFAMsi TIGR01502. B_methylAsp_ase. 1 hit.
    ProtoNeti Search...

    Publicationsi

    1. "Cloning, sequencing, and expression in Escherichia coli of the Clostridium tetanomorphum gene encoding beta-methylaspartase and characterization of the recombinant protein."
      Goda S.K., Minton N.P., Botting N.P., Gani D.
      Biochemistry 31:10747-10756(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-26, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.
      Strain: ATCC 15920 / DSM 528 / H1.
    2. "Cloning and sequencing of glutamate mutase component E from Clostridium tetanomorphum."
      Brecht M., Kellermann J., Plueckthun A.
      FEBS Lett. 319:84-89(1993) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-24, PROTEIN SEQUENCE OF 1-24.
      Strain: ATCC 15920 / DSM 528 / H1.
    3. "Cloning and sequencing of glutamate mutase component E from Clostridium tetanomorphum. Organization of the mut genes."
      Holloway D.E., Marsh E.N.G.
      FEBS Lett. 317:44-48(1993) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-71.
      Strain: NCIMB 11547.
    4. "Cloning and sequencing of glutamate mutase component S from Clostridium tetanomorphum. Homologies with other cobalamin-dependent enzymes."
      Marsh E.N.G., Holloway D.E.
      FEBS Lett. 310:167-170(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 1-15.
      Strain: NCIMB 11547.
    5. "The purification and properties of beta-methylaspartase."
      Barker H.A., Smyth R.D., Wilson R.M., Weissbach H.
      J. Biol. Chem. 234:320-328(1959) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, SUBSTRATE SPECIFICITY, COFACTOR.
      Strain: ATCC 15920 / DSM 528 / H1.
    6. "Alteration of the diastereoselectivity of 3-methylaspartate ammonia lyase by using structure-based mutagenesis."
      Raj H., Weiner B., Veetil V.P., Reis C.R., Quax W.J., Janssen D.B., Feringa B.L., Poelarends G.J.
      ChemBioChem 10:2236-2245(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF HIS-194; GLN-329 AND LYS-331, SUBSTRATE SPECIFICITY, ACTIVE SITE, SUBUNIT.
      Strain: ATCC 15920 / DSM 528 / H1.
    7. "The structure of 3-methylaspartase from Clostridium tetanomorphum functions via the common enolase chemical step."
      Asuncion M., Blankenfeldt W., Barlow J.N., Gani D., Naismith J.H.
      J. Biol. Chem. 277:8306-8311(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, ACTIVE SITE, COFACTOR, SUBUNIT.
    8. Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS AND MAGNESIUM, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF GLN-73 AND LEU-384, COFACTOR, SUBUNIT.

    Entry informationi

    Entry nameiMAAL_CLOTT
    AccessioniPrimary (citable) accession number: Q05514
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: February 1, 1995
    Last sequence update: February 1, 1995
    Last modified: October 1, 2014
    This is version 75 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Direct protein sequencing

    Documents

    1. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    2. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    3. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3