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Q05514

- MAAL_CLOTT

UniProt

Q05514 - MAAL_CLOTT

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Protein

Methylaspartate ammonia-lyase

Gene
N/A
Organism
Clostridium tetanomorphum
Status
Reviewed - Annotation score: 5 out of 5 - Experimental evidence at protein leveli

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

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)

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)

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)

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)

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)

pH dependencei

Optimum pH is 9.7.

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.

Pathwayi

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei172 – 1721L-threo-beta-methylaspartate By similarity
Sitei194 – 1941Transition state stabilizer
Metal bindingi238 – 2381Magnesium
Metal bindingi273 – 2731Magnesium
Metal bindingi307 – 3071Magnesium
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
Complete GO annotation...

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.

Miscellaneous databases

EvolutionaryTraceiQ05514.

Family & Domainsi

Region

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

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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: July 9, 2014
This is version 74 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

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