ADSL » Adenylosuccinate lyase (ASL)

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Protein also known as: Adenylosuccinase (ASase).

Gene name: ADSL

Family name: Lyase 1 » Adenylosuccinate lyase

One or more isoforms of this protein have been shown to exist at protein level

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1 39 2

GENE REF ISO

Function

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GO molecular function

(S)-2-(5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamido)succinate AMP-lyase (fumarate-forming) activitydefinition[GO:0070626]

IEAEC 2 GO

Isoform Iso 1 but not isoform Iso 2 N6-(1,2-dicarboxyethyl)AMP AMP-lyase (fumarate-forming) activitydefinition[GO:0004018]

Evidence 1: Inferred from Direct Assay UniProtKB

Expression, purification, and characterization of stable, recombinant human adenylosuccinate lyase.

Lee P., Colman R.F.

Protein Expr. Purif. 51, 227-234 (2007) [Full text:10.1016/j.pep.2006.07.023] [PubMed:16973378]

The full length human adenylosuccinate lyase gene was generated by a PCR method using a plasmid encoding a truncated human enzyme as template, and was cloned into a pET-14b vector. Human adenylosuccinate lyase was overexpressed in Escherichia coli Rosetta 2(DE3)pLysS as an N-terminal histidine-tagged protein and was purified to homogeneity by a nickel-nitriloacetic acid column at room temperature. The histidine tag was removed from the human enzyme by thrombin digestion and the adenylosuccinate lyase was purified by Sephadex G-100 gel filtration. The histidine-tagged and non-tagged adenylosuccinate lyases exhibit similar values of Vmax and Km for S-AMP. Analytical ultracentrifugation and circular dichroism revealed, respectively, that the histidine-tagged enzyme is in tetrameric form with a molecular weight of 220 kDa and contains predominantly alpha-helical structure. This is the first purification procedure to yield a stable form of human adenylosuccinate lyase. The enzyme is stable for at least 5 days at 25 degrees C, and upon rapid freezing and thawing. Temperature as well as reducing agent (DTT) play critical roles in determining the stability of the human adenylosuccinate lyase.
Evidence 2: Inferred from Direct Assay UniProtKB

Human adenylosuccinate lyase (ADSL), cloning and characterization of full-length cDNA and its isoform, gene structure and molecular basis for ADSL deficiency in six patients.

Kmoch S., Hartmannová H., Stibůrková B., Krijt J., Zikánová M., Sebesta I.

Hum. Mol. Genet. 9, 1501-1513 (2000) [PubMed:10888601]

Adenylosuccinate lyase (ADSL) is a bifunctional enzyme acting in de novo purine synthesis and purine nucleotide recycling. ADSL deficiency is a selectively neuronopathic disorder with psychomotor retardation and epilepsy as leading traits. Both dephosphorylated enzyme substrates, succinylaminoimidazole-carboxamide riboside (SAICAr) and succinyladenosine (S-Ado), accumulate in the cerebrospinal fluid (CSF) of affected individuals with S-Ado/SAICAr concentration ratios proportional to the phenotype severity. We studied the disorder at various levels in a group of six patients with ADSL deficiency. We identified the complete ADSL cDNA and its alternatively spliced isoform resulting from exon 12 skipping. Both mRNA isoforms were expressed in all the tissues studied with the non-spliced form 10-fold more abundant. Both cDNAs were expressed in Escherichia coli and functionally characterized at the protein level. The results showed only the unspliced ADSL to be active. The gene consists of 13 exons spanning 23 kb. The promotor region shows typical features of the housekeeping gene. Eight mutations were identified in a group of six patients. The expression studies of the mutant proteins carried out in an attempt to study genotype-phenotype correlation showed that the level of residual enzyme activity correlates with the severity of the clinical phenotype. All the mutant enzymes studied in vitro displayed a proportional decrease in activity against both of their substrates. However, this was not concordant with strikingly different concentration ratios in the CSF of individual patients. This suggests either different in vivo enzyme activities against each of the substrates and/or their different turnover across the CSF-blood barrier, which may be decisive in determining disease severity.

refs

IDAUniProtKB

GO biological process

'de novo' AMP biosynthetic processdefinition[GO:0044208]

IEAUniPathway

'de novo' IMP biosynthetic processdefinition[GO:0006189]

IEAUniPathway

Aerobic respirationdefinition[GO:0009060] ‹silver

IEAOrtholog Compara

AMP biosynthetic processdefinition[GO:0006167]

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IDAUniProtKB

Protein tetramerizationdefinition[GO:0051262]

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IDAUniProtKB

Purine nucleotide biosynthetic processdefinition[GO:0006164]

Evidence 1: Inferred by Curator UniProtKB

Human adenylosuccinate lyase (ADSL), cloning and characterization of full-length cDNA and its isoform, gene structure and molecular basis for ADSL deficiency in six patients.

Kmoch S., Hartmannová H., Stibůrková B., Krijt J., Zikánová M., Sebesta I.

Hum. Mol. Genet. 9, 1501-1513 (2000) [PubMed:10888601]

Adenylosuccinate lyase (ADSL) is a bifunctional enzyme acting in de novo purine synthesis and purine nucleotide recycling. ADSL deficiency is a selectively neuronopathic disorder with psychomotor retardation and epilepsy as leading traits. Both dephosphorylated enzyme substrates, succinylaminoimidazole-carboxamide riboside (SAICAr) and succinyladenosine (S-Ado), accumulate in the cerebrospinal fluid (CSF) of affected individuals with S-Ado/SAICAr concentration ratios proportional to the phenotype severity. We studied the disorder at various levels in a group of six patients with ADSL deficiency. We identified the complete ADSL cDNA and its alternatively spliced isoform resulting from exon 12 skipping. Both mRNA isoforms were expressed in all the tissues studied with the non-spliced form 10-fold more abundant. Both cDNAs were expressed in Escherichia coli and functionally characterized at the protein level. The results showed only the unspliced ADSL to be active. The gene consists of 13 exons spanning 23 kb. The promotor region shows typical features of the housekeeping gene. Eight mutations were identified in a group of six patients. The expression studies of the mutant proteins carried out in an attempt to study genotype-phenotype correlation showed that the level of residual enzyme activity correlates with the severity of the clinical phenotype. All the mutant enzymes studied in vitro displayed a proportional decrease in activity against both of their substrates. However, this was not concordant with strikingly different concentration ratios in the CSF of individual patients. This suggests either different in vivo enzyme activities against each of the substrates and/or their different turnover across the CSF-blood barrier, which may be decisive in determining disease severity.

ref

ICUniProtKB

Response to hypoxiadefinition[GO:0001666] ‹silver

IEAOrtholog Compara

Response to muscle activitydefinition[GO:0014850] ‹silver

IEAOrtholog Compara

Response to nutrientdefinition[GO:0007584] ‹silver

IEAOrtholog Compara

Response to starvationdefinition[GO:0042594] ‹silver

IEAOrtholog Compara

Enzymatic activity

This protein acts as an enzyme. It is known to catalyze the following reactions

N(6)-(1,2-dicarboxyethyl)AMP ⇄ fumarate + AMP.

CuratedUniProtKB

(S)-2-(5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamido)succinate ⇄ fumarate + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide.

CuratedUniProtKB

Pathways

This protein is involved in the following pathways

Purine metabolism; AMP biosynthesis via de novo pathway; AMP from IMP: step 2/2

CuratedUniProtKB

Purine metabolism; IMP biosynthesis via de novo pathway; 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxamide from 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate: step 2/2

CuratedUniProtKB

According to KEGG, this protein belongs to the following pathways:

Alanine, aspartate and glutamate metabolism hsa00250+158

Metabolic pathways hsa01100+158

Purine metabolism hsa00230+158

According to Reactome, this protein belongs to the following pathway:

Metabolism REACT_111217

Keywords

Biological process

Purine biosynthesis definition [KW-0658]

Molecular function

Lyase definition [KW-0456]

Technical term

Reference proteome definition [KW-1185]

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Function

Keywords

neXtProt