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Literature summary for 4.3.2.2 extracted from

  • De Zoysa Ariyananda, L.; Antonopoulos, C.; Currier, J.; Colman, R.F.
    In vitro hybridization and separation of hybrids of human adenylosuccinate lyase from wild-type and disease-associated mutant enzymes (2011), Biochemistry, 50, 1336-1346.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.3.2.2

Application

Application Comment Organism
medicine generation of in vitro hybrids that mimic compound heterozygote ASL for studies on disease-associated mutant enzymes. His-tagged wild-type/non-His-tagged wild-type, His-tagged wild-type/non-His-tagged R396C, His-tagged wild-type/non-His-tagged R396H, His-tagged R194C/non-His-tagged R396C, and His-tagged L311V/non-His-tagged R396H enzyme pairs are used in various hybrids generated by denaturing pairs of enzymes in 1 M guanidinium chloride and renaturing them by removing the denaturant. The hybrids have predominant amounts of heterotetramers. Analysis of the Vmax values of the hybrids indicates that most of the subunits behave independently. However, the hybrid tetramers retain weak positive cooperativity Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
-
 Homo sapiens

Protein Variants

Protein Variants Comment Organism
L311V about 90% of wild-type actitivy Homo sapiens
R194C about 100% of wild-type actitivy Homo sapiens
R396C about 20% of wild-type actitivy Homo sapiens
R396H about 10% of wild-type actitivy Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57000
-
 4 * 57000, calculated, His-tagged protein Homo sapiens
228000
-
 gel filtration, His-tagged protein Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1
-
 mutant R396H, pH 7.4, 25°C Homo sapiens
1.1
-
 mutant R396H, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens
1.6
-
 mutant R396C, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens
1.8
-
 mutant R396C, pH 7.4, 25°C Homo sapiens
7.5
-
 His-tagged mutant L311V, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens
7.8
-
 His-tagged mutant L311V, pH 7.4, 25°C Homo sapiens
8.3
-
 His-tagged wild-type, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens
8.4
-
 wild-type, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens
8.5
-
 His-tagged wild-type, pH 7.4, 25°C Homo sapiens
8.6
-
 wild-type, pH 7.4, 25°C Homo sapiens
8.7
-
 His-tagged mutant R194C, pH 7.4, 25°C Homo sapiens
8.9
-
 His-tagged mutant R194C, after treatment with guanidiniumHCl, pH 7.4, 25°C Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(Nomega-L-arginino)succinate
-
 Homo sapiens fumarate + L-arginine
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 57000, calculated, His-tagged protein Homo sapiens