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Literature summary for 3.2.1.68 extracted from

  • Sundberg, M.; Pfister, B.; Fulton, D.; Bischof, S.; Delatte, T.; Eicke, S.; Stettler, M.; Smith, S.M.; Streb, S.; Zeeman, S.C.
    The heteromultimeric debranching enzyme involved in starch synthesis in Arabidopsis requires both isoamylase1 and isoamylase2 subunits for complex stability and activity (2013), PLoS ONE, 8, e75223.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.68

Application

Application Comment Organism
analysis isoamylase-type debranching enzymes (ISAs) play an important role in determining starch structure Arabidopsis thaliana

Cloned(Commentary)

Cloned (Comment) Organism
coexpression of wild-type His-tagged ISA1 and ISA2, and of mutant D367A ISA1 with wild-type ISA2, in Escherichia coli strain BL21(DE3) Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
D367A site-directed mutagenesis of the catalytic nucleophile Asp-367 in ISA1, the mutation abolishes the catalytic activity of the enzyme complex Arabidopsis thaliana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
 comparable mobility in the native PAGE of native enzyme recombinnat enzyme complex Arabidopsis thaliana
150000
-
 x * 150000, ISA1 and ISA2 as stable dimer, SDS-PAGE Arabidopsis thaliana
450000
-
 native ISA1-ISA2 complex, gel filtration Arabidopsis thaliana
750000
-
 recombinant ISA1-ISA2 complex, gel filtration Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Arabidopsis thaliana synthesis of amylopectin and phytoglycogen by associated and separated ISA1 and ISA2, respectively, overview ?
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana O04196 isozyme ISA1; genes isa1 and isa2
-
Arabidopsis thaliana Q8L735 isozyme ISA2; genes isa1 and isa2
-

Purification (Commentary)

Purification (Comment) Organism
copurification of native isozymes ISA1 and ISA2 partially from leaves by gel filtration, copurification of TAP-tagged ISA1 and ISA2 from Escherichia coli strain BL21(DE3) by tandem-affinity chromatography on TEV and calmodulin, followed by gel filtration Arabidopsis thaliana

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
 Arabidopsis thaliana
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
maize beta-limit dextrin + H2O
-
 Arabidopsis thaliana ?
-
 ?
additional information synthesis of amylopectin and phytoglycogen by associated and separated ISA1 and ISA2, respectively, overview Arabidopsis thaliana ?
-
 ?
oyster glycogen + H2O
-
 Arabidopsis thaliana ?
-
 ?
potato amylopectin + H2O
-
 Arabidopsis thaliana ?
-
 ?

Subunits

Subunits Comment Organism
? x * 150000, ISA1 and ISA2 as stable dimer, SDS-PAGE Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
ISA
-
 Arabidopsis thaliana
ISA1
-
 Arabidopsis thaliana
ISA2
-
 Arabidopsis thaliana
isoamylase-type debranching enzyme
-
 Arabidopsis thaliana

General Information

General Information Comment Organism
malfunction loss of isozyme ISA1 or ISA2 causes phytoglycogen accumulation Arabidopsis thaliana
additional information isozyme ISA1 interacts with its homologue ISA2, but no evidence for interaction with other starch biosynthetic enzymes. ISA1 and ISA2 form a heteromultimeric enzyme with ISA1 being the catalytic subunit and ISA2 subunit not catalytically active Arabidopsis thaliana
physiological function the ISA1 class of DBE is the one primarily associated with amylopectin synthesis Arabidopsis thaliana