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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

Mg2+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
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 – 1110Combined sources
Beta strandi14 – 185Combined sources
Helixi20 – 245Combined sources
Beta strandi28 – 303Combined sources
Beta strandi33 – 364Combined sources
Beta strandi44 – 496Combined sources
Beta strandi52 – 598Combined sources
Beta strandi64 – 696Combined sources
Turni73 – 764Combined sources
Helixi86 – 9611Combined sources
Helixi98 – 1014Combined sources
Helixi109 – 11810Combined sources
Helixi128 – 14619Combined sources
Helixi150 – 1589Combined sources
Helixi179 – 1868Combined sources
Beta strandi190 – 1945Combined sources
Helixi200 – 2045Combined sources
Helixi209 – 22517Combined sources
Beta strandi234 – 2385Combined sources
Helixi242 – 2465Combined sources
Turni247 – 2493Combined sources
Helixi251 – 26515Combined sources
Beta strandi270 – 2734Combined sources
Helixi281 – 29818Combined sources
Beta strandi302 – 3065Combined sources
Helixi313 – 3219Combined sources
Beta strandi325 – 3306Combined sources
Helixi333 – 3353Combined sources
Helixi339 – 35012Combined sources
Beta strandi354 – 3574Combined sources
Helixi365 – 37814Combined sources
Beta strandi381 – 3844Combined sources
Beta strandi387 – 3915Combined sources
Helixi392 – 41019Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
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/MAL.
[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

        10         20         30         40         50
MKIVDVLCTP GLTGFYFDDQ RAIKKGAGHD GFTYTGSTVT EGFTQVRQKG
60 70 80 90 100
ESISVLLVLE DGQVAHGDCA AVQYSGAGGR DPLFLAKDFI PVIEKEIAPK
110 120 130 140 150
LIGREITNFK PMAEEFDKMT VNGNRLHTAI RYGITQAILD AVAKTRKVTM
160 170 180 190 200
AEVIRDEYNP GAEINAVPVF AQSGDDRYDN VDKMIIKEAD VLPHALINNV
210 220 230 240 250
EEKLGLKGEK LLEYVKWLRD RIIKLRVRED YAPIFHIDVY GTIGAAFDVD
260 270 280 290 300
IKAMADYIQT LAEAAKPFHL RIEGPMDVED RQKQMEAMRD LRAELDGRGV
310 320 330 340 350
DAELVADEWC NTVEDVKFFT DNKAGHMVQI KTPDLGGVNN IADAIMYCKA
360 370 380 390 400
NGMGAYCGGT CNETNRSAEV TTNIGMACGA RQVLAKPGMG VDEGMMIVKN
410
EMNRVLALVG RRK
Length:413
Mass (Da):45,534
Last modified:February 1, 1995 - v1
Checksum:i4451923DB035EF13
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
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:
EMBLi
GenBanki
DDBJi
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:
PDBei
RCSB PDBi
PDBji
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/MAL.
[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: November 26, 2014
This is version 77 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