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Protein

Methylaspartate ammonia-lyase

Gene
N/A
Organism
Clostridium tetanomorphum
Status
Reviewed-Annotation score: 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
  1. 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
  2. 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
  3. 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
  4. 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
  5. 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: L-glutamate degradation via mesaconate pathway

This protein is involved in step 2 of the subpathway that synthesizes acetate and pyruvate from L-glutamate.
Proteins known to be involved in the 4 steps of the subpathway in this organism are:
  1. Glutamate mutase sigma subunit (glmS), Glutamate mutase epsilon subunit (glmE)
  2. Methylaspartate ammonia-lyase
  3. no protein annotated in this organism
  4. no protein annotated in this organism
This subpathway is part of the pathway L-glutamate degradation via mesaconate pathway, which is itself part of Amino-acid degradation.
View all proteins of this organism that are known to be involved in the subpathway that synthesizes acetate and pyruvate from L-glutamate, the pathway L-glutamate degradation via mesaconate pathway and in Amino-acid degradation.

Sites

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Binding sitei172L-threo-beta-methylaspartateBy similarity1
Sitei194Transition state stabilizer1
Metal bindingi238Magnesium2 Publications1
Metal bindingi273Magnesium2 Publications1
Metal bindingi307Magnesium2 Publications1
Binding sitei329L-threo-beta-methylaspartate1
Active sitei331Proton acceptor2 Publications1
Binding sitei361L-threo-beta-methylaspartate1

GO - Molecular functioni

GO - Biological processi

Complete GO annotation...

Keywords - Molecular functioni

Lyase

Keywords - Ligandi

Cobalamin, Cobalt, Magnesium, Metal-binding

Enzyme and pathway databases

BioCyciMetaCyc:MONOMER-1103.
BRENDAi4.3.1.2. 1527.
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)DescriptionActionsGraphical viewLength
Mutagenesisi73Q → 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 Publication1
Mutagenesisi194H → 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 Publication1
Mutagenesisi194H → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi329Q → 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 Publication1
Mutagenesisi329Q → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi331K → A: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi331K → G: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi331K → H: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi331K → Q: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi331K → R: It abolishes deaminase and aminase activities and does not show any major conformational changes. 1 Publication1
Mutagenesisi384L → A: It has very broad electrophile scope and excellent regio- and enantioselectivity in the amination reaction. 1 Publication1

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
ChainiPRO_00000845471 – 413Methylaspartate ammonia-lyaseAdd BLAST413

Interactioni

Subunit structurei

Homodimer.4 Publications

Structurei

Secondary structure

1413
Legend: HelixTurnBeta strandPDB Structure known for this area
Show more details
Feature keyPosition(s)DescriptionActionsGraphical viewLength
Beta strandi2 – 11Combined sources10
Beta strandi14 – 18Combined sources5
Helixi20 – 24Combined sources5
Beta strandi28 – 30Combined sources3
Beta strandi33 – 36Combined sources4
Beta strandi44 – 49Combined sources6
Beta strandi52 – 59Combined sources8
Beta strandi64 – 69Combined sources6
Turni73 – 76Combined sources4
Helixi86 – 96Combined sources11
Helixi98 – 101Combined sources4
Helixi109 – 118Combined sources10
Helixi128 – 146Combined sources19
Helixi150 – 158Combined sources9
Helixi179 – 186Combined sources8
Beta strandi190 – 194Combined sources5
Helixi200 – 204Combined sources5
Helixi209 – 225Combined sources17
Beta strandi234 – 238Combined sources5
Helixi242 – 246Combined sources5
Turni247 – 249Combined sources3
Helixi251 – 265Combined sources15
Beta strandi270 – 273Combined sources4
Helixi281 – 298Combined sources18
Beta strandi302 – 306Combined sources5
Helixi313 – 321Combined sources9
Beta strandi325 – 330Combined sources6
Helixi333 – 335Combined sources3
Helixi339 – 350Combined sources12
Beta strandi354 – 357Combined sources4
Helixi365 – 378Combined sources14
Beta strandi381 – 384Combined sources4
Beta strandi387 – 391Combined sources5
Helixi392 – 410Combined sources19

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB 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.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiQ05514.

Family & Domainsi

Region

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Regioni360 – 361L-threo-beta-methylaspartate bindingBy similarity2

Sequence similaritiesi

Family and domain databases

CDDicd03314. MAL. 1 hit.
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. 2 hits.
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.
PIRiB44285.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB 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.
ModBaseiSearch...
MobiDBiSearch...

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Enzyme and pathway databases

UniPathwayiUPA00561; UER00618.
BioCyciMetaCyc:MONOMER-1103.
BRENDAi4.3.1.2. 1527.

Miscellaneous databases

EvolutionaryTraceiQ05514.

Family and domain databases

CDDicd03314. MAL. 1 hit.
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. 2 hits.
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.
ProtoNetiSearch...

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 2, 2016
This is version 85 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

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.