Literature summary for 3.4.24.15 extracted from

Westlake, T.J.; Ricci, W.A.; Popescu, G.V.; Popescu, S.C.
Dimerization and thiol sensitivity of the salicylic acid binding thimet oligopeptidases TOP1 and TOP2 define their functions in redox-sensitive cellular pathways (2015), Front. Plant Sci., 6, 327 .

Search for more literature for 3.4.24.15

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged enzyme TOP1 in Escherichia coli strain Bl21. Arabidopsis protoplasts are prepared and co-transformed with pairs of plasmid constructs containing the coding sequences of TOP1 or TOP2, cloned in frame with either the N-terminal or the C-terminal halves of the Renilla luciferase coding sequence (TOP1-orTOP2-NLuc and TOP1- or TOP2- CLuc). Interactions between TOPs-Luc fusions are detected by measuring luminescence released upon the restoration of luciferase enzymatic activity. Protoplasts expressing luciferase terminal fusions with known interactors MKK5 and HOPF2	Arabidopsis thaliana
recombinant expression of His-tagged enzyme TOP2 in Escherichia coli strain Bl21. Arabidopsis protoplasts are prepared and co-transformed with pairs of plasmid constructs containing the coding sequences of TOP1 or TOP2, cloned in frame with either the N-terminal or the C-terminal halves of the Renilla luciferase coding sequence (TOP1-orTOP2-NLuc and TOP1- or TOP2- CLuc). Interactions between TOPs-Luc fusions are detected by measuring luminescence released upon the restoration of luciferase enzymatic activity. Protoplasts expressing luciferase terminal fusions with known interactors MKK5 and HOPF2	Arabidopsis thaliana

Cloned (Comment)

Organism

recombinant expression of His-tagged enzyme TOP1 in Escherichia coli strain Bl21. Arabidopsis protoplasts are prepared and co-transformed with pairs of plasmid constructs containing the coding sequences of TOP1 or TOP2, cloned in frame with either the N-terminal or the C-terminal halves of the Renilla luciferase coding sequence (TOP1-orTOP2-NLuc and TOP1- or TOP2- CLuc). Interactions between TOPs-Luc fusions are detected by measuring luminescence released upon the restoration of luciferase enzymatic activity. Protoplasts expressing luciferase terminal fusions with known interactors MKK5 and HOPF2

Arabidopsis thaliana

recombinant expression of His-tagged enzyme TOP2 in Escherichia coli strain Bl21. Arabidopsis protoplasts are prepared and co-transformed with pairs of plasmid constructs containing the coding sequences of TOP1 or TOP2, cloned in frame with either the N-terminal or the C-terminal halves of the Renilla luciferase coding sequence (TOP1-orTOP2-NLuc and TOP1- or TOP2- CLuc). Interactions between TOPs-Luc fusions are detected by measuring luminescence released upon the restoration of luciferase enzymatic activity. Protoplasts expressing luciferase terminal fusions with known interactors MKK5 and HOPF2

Arabidopsis thaliana

Inhibitors

Inhibitors	Comment	Organism	Structure
1,4-dithiothreitol	DTT inhibits both, TOP1 and TOP2. As with TOP1, both TOP2 monomeric and dimeric fractions are inhibited by the thiol-based reductant; DTT inhibits both, TOP1 and TOP2. As with TOP1, both TOP2 monomeric and dimeric fractions are inhibited by the thiol-based reductant	Arabidopsis thaliana

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	-	-	-
Arabidopsis thaliana	A0A1R7T3A4	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme TOP1 from Escherichia coli strain Bl21 by cobalt affinity chromatography and gel filtration	Arabidopsis thaliana
recombinant His-tagged enzyme TOP2 from Escherichia coli strain Bl21 by cobalt affinity chromatography and gel filtration	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys (Dnp)-OH + H2O	-	Arabidopsis thaliana	?	-	?

Subunits

Subunits	Comment	Organism
dimer	TOP1 and TOP2 assemble in homo- and heterodimers. And TOP1 and TOP2 form transient monomers and dimers. Salicylic acid and dithiothreitol influence the dimer-monomer balance of TOP1 and TOP2	Arabidopsis thaliana

Synonyms

Synonyms	Comment	Organism
Top1	-	Arabidopsis thaliana
Top2	-	Arabidopsis thaliana

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	8.5	broad optimum	Arabidopsis thaliana

General Information

General Information	Comment	Organism
metabolism	dimerization and thiol sensitivity of the salicylic acid binding thimet oligopeptidases TOP1 and TOP2 define their functions in redox-sensitive cellular pathways. The interplay between TOP1-controlled chloroplastic events and the cytosolic TOP2 modulates the development of programmed cell death through the ROS-SA-AOX axis. TOP1 and TOP2 mediate plant responses to oxidative stress through spatially separated pathways and positions proteolysis in a network for plant response to diverse stressors, overview	Arabidopsis thaliana
additional information	molecular dynamics simulations and modelling	Arabidopsis thaliana
physiological function	TOP1 and TOP2 interact with themselves and each other and their ability to associate in dimers is influenced by salicylic acid and the thiol-based reductant DTT. The interplay between TOP1-controlled chloroplastic events and the cytosolic TOP2 modulates the development of programmed cell death through the ROS-SA-AOX axis. TOP1 and TOP2 are modulators of the plant tolerance to methyl viologen (MV), and exogenous salicylic acid alleviates the toxicity of MV in top background. TOP1 and TOP2 mediate plant responses to oxidative stress through spatially separated pathways and positions proteolysis in a network for plant response to diverse stressors, overview. TOP1 and TOP2 specifically mediate plant responses to MV during early development via, at least partially, distinct pathways. Also, it suggest that TOPs do not have broad, unspecific roles in mediating plant's oxidative stress response	Arabidopsis thaliana