Cloned (Comment) | Organism |
---|---|
gene isa1, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3), co-expression of untagged ISA1 subunit with C-terminally His8-tagged subunit ISA2 in Escherichia coli | Arabidopsis thaliana |
gene isa2, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3) or co-expression with untagged ISA1 subunit in Escherichia coli | Zea mays |
gene isa2, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3) or co-expression with untagged ISA1 subunit in Escherichia coli | Arabidopsis thaliana |
gene su-1, single expression of C-terminally His8-tagged subunit ISA1 in Escherichia coli strain BL21(DE3), co-expression of untagged ISA1 subunit with C-terminally His8-tagged subunit ISA2 in Escherichia coli. Zea mays ISA1 subunit is heterologously expressed in Arabidopsis thaliana lacking endogenous ISA1 or lacking both native ISA1 and ISA2 via transfection with Agrobacterium tumefaciens strain GV3101. The maize protein functions in Arabidopsis leaves to support nearly normal starch metabolism in the absence of any native ISA1 or ISA2 | Zea mays |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a single mutant line isa2-2 by T-DNA insertion mutagenesis. The isa1-1 isa2-2 double mutant line is obtained by a cross between the two corresponding single mutants and screening of the progeny by PCR amplification of genomic DNA using a T-DNA end primer and gene-specific primers. The phenotype of the isa1-1 isa2-2 double mutant is complemented but the ZmISA1 activity | Arabidopsis thaliana |
additional information | construction of single mutant line isa1-1 by T-DNA insertion mutagenesis. The isa1-1 isa2-2 double mutant line is obtained by a cross between the two corresponding single mutants and screening of the progeny by PCR amplification of genomic DNA using a T-DNA end primer and gene-specific primers. The phenotype of the isa1-1 isa2-2 double mutant is complemented but the ZmISA1 activity, Zea mays ISA1 subunit is expressed via transfection with Agrobacterium tumefaciens strain GV3101 in Arabidopsis thaliana lacking endogenous ISA1 or lacking both native ISA1 and ISA2. The maize protein functions in Arabidopsis leaves to support nearly normal starch metabolism in the absence of any native ISA1 or ISA2 | Arabidopsis thaliana |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Zea mays | 9507 | - |
chloroplast | - |
Arabidopsis thaliana | 9507 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
85000 | - |
1 * 85000, ISA1 monomer, SDS-PAGE | Zea mays |
85000 | - |
2 * 85000, ISA1 dimer, SDS-PAGE | Zea mays |
158000 | - |
ISA2 dimer, gel filtration and analytical ultracentrifugation | Zea mays |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
amylopectin + H2O | Arabidopsis thaliana | - |
? | - |
? | |
amylopectin + H2O | Zea mays | - |
? | - |
? | |
amylopectin + H2O | Arabidopsis thaliana Col-0 | - |
? | - |
? | |
Arabidopsis starch + H2O | Arabidopsis thaliana | - |
? | - |
? | |
Arabidopsis starch + H2O | Arabidopsis thaliana Col-0 | - |
? | - |
? | |
beta-limit dextrin + H2O | Arabidopsis thaliana | - |
? | - |
? | |
beta-limit dextrin + H2O | Arabidopsis thaliana Col-0 | - |
? | - |
? | |
maize starch + H2O | Zea mays | - |
? | - |
? | |
additional information | Zea mays | Zea mays subunit ISA 2 alone is catalytically inactive | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | O04196 | gene isa1 | - |
Arabidopsis thaliana | Q8L735 | gene isa2 | - |
Arabidopsis thaliana Col-0 | O04196 | gene isa1 | - |
Arabidopsis thaliana Col-0 | Q8L735 | gene isa2 | - |
Zea mays | - |
gene isa2 | - |
Zea mays | B6U0X5 | gene su-1 | - |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His8-tagged single subunit ISA1 or ISA1 in complex with His8-tagged ISA2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Arabidopsis thaliana |
recombinant C-terminally His8-tagged single subunit ISA1 or ISA1 in complex with His8-tagged ISA2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Zea mays |
recombinant C-terminally His8-tagged single subunit ISA2 or His8-tagged ISA2 in complex with ISA1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Zea mays |
recombinant C-terminally His8-tagged single subunit ISA2 or His8-tagged ISA2 in complex with ISA1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Arabidopsis thaliana |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
endosperm | only ISA1, no ISA2 | Zea mays | - |
leaf | - |
Arabidopsis thaliana | - |
leaf | ISA1 and ISA2 | Zea mays | - |
additional information | not in endosperm | Zea mays | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
amylopectin + H2O | - |
Arabidopsis thaliana | ? | - |
? | |
amylopectin + H2O | - |
Zea mays | ? | - |
? | |
amylopectin + H2O | - |
Arabidopsis thaliana Col-0 | ? | - |
? | |
Arabidopsis starch + H2O | - |
Arabidopsis thaliana | ? | - |
? | |
Arabidopsis starch + H2O | - |
Arabidopsis thaliana Col-0 | ? | - |
? | |
beta-limit dextrin + H2O | - |
Arabidopsis thaliana | ? | - |
? | |
beta-limit dextrin + H2O | - |
Zea mays | ? | - |
? | |
beta-limit dextrin + H2O | - |
Arabidopsis thaliana Col-0 | ? | - |
? | |
maize starch + H2O | - |
Zea mays | ? | - |
? | |
additional information | Zea mays subunit ISA 2 alone is catalytically inactive | Zea mays | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 85000, ISA1 dimer, SDS-PAGE | Zea mays |
monomer | 1 * 85000, ISA1 monomer, SDS-PAGE | Zea mays |
Synonyms | Comment | Organism |
---|---|---|
AtISA1 | - |
Arabidopsis thaliana |
AtISA2 | - |
Arabidopsis thaliana |
ISA1 | - |
Arabidopsis thaliana |
ISA1 | - |
Zea mays |
ISA2 | - |
Zea mays |
ISA2 | - |
Arabidopsis thaliana |
isoamylase-type starch debranching enzyme 1 | - |
Arabidopsis thaliana |
isoamylase-type starch debranching enzyme 1 | - |
Zea mays |
isoamylase-type starch debranching enzyme 2 | - |
Zea mays |
isoamylase-type starch debranching enzyme 2 | - |
Arabidopsis thaliana |
ZmISA1 | - |
Zea mays |
ZmISA2 | - |
Zea mays |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycosyl hydrolase family 13 GH13, and harbors a carbohydrate-binding module family 48 (CBM48) domain | Zea mays |
additional information | Arabidopsis thaliana subunit ISA1 requires subunit ISA2 as a partner for enzymatic function. Images of purified starch granules of wild-type and mutant lines, overview | Arabidopsis thaliana |
additional information | Arabidopsis thaliana subunit ISA1 requires subunit ISA2 as a partner for enzymatic function. Images of purified starch granules of wild-type and mutant lines, overview. Recombinant AtISA1 is capable of enzymatic activity if mixed with recombinant AtISA2 but does not display such function on its own | Arabidopsis thaliana |
additional information | Zea mays subunit ISA1 is active by itself and does not require subunit ISA2 for activity, subunit ISA 2 alone is catalytically inactive | Zea mays |
additional information | Zea mays subunit ISA1 is active by itself, either in vitro or in transgenic Arabidopsis leaves, and does not require subunit ISA2 for activity | Zea mays |