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5.6.1.1: microtubule-severing ATPase

This is an abbreviated version!
For detailed information about microtubule-severing ATPase, go to the full flat file.

Word Map on EC 5.6.1.1

Reaction

n ATP + n H2O +

a microtubule
= n ADP + n phosphate + (n+1) alpha/beta tubulin heterodimers

Synonyms

adseverin, Arabidopsis thaliana katanin small subunit, AtKSS, D-spastin, DGL1, Dm-Kat60, Drosophila katanin Dm-Kat60, EC 3.6.4.3, kananin, Kat60, katanin, KATANIN 1, katanin p60, katanin p60 ATPase-containing subunit A1, katanin p60 microtubule-severing protein, KATNAL1, kinesin spindle protein, kp60-NTD, KSP, mei-1, MEI-1/MEI-2 complex, MEI-1/MEI-2 katanin complex, microtuble severing AAA ATPase spastin, microtuble-severing AAA ATPase, microtuble-severing ATPase spastin, microtubule severing protein, microtubule-severing AAA ATPase, microtubule-severing ATPase, microtubule-severing complex katanin, microtubule-severing complex MEI-1/MEI-2 katanin, microtubule-severing protein, microtubule-stimulated ATPase, MT-severing AAA ATPase, P60, p60 KAT1p, p60 katanin, P60-kananin, P60-katanin, p81-p60, Pf15, plant microtubule-severing protein, rice KTN1, scinderin, SPAS-1, spastin, spastin AAA domain, SPG4, Sug1p, vertebrate katanin

ECTree

     5 Isomerases
         5.6 Isomerases altering macromolecular conformation
             5.6.1 Enzymes altering polypeptide conformation or assembly
                5.6.1.1 microtubule-severing ATPase

Purification

Purification on EC 5.6.1.1 - microtubule-severing ATPase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
90% pure after Ni2+-NTA agarose, analyzed by SDS-PAGE
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baculovirus-expressed, recombinant DM-Kat60
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copurification of p81 and p60
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GSH-Sepharose affinity chromatography, ion exchange chromatography, cleavage with precission protease, GSH-Sepharose affinity chromatography and gel filtration
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GST tagged p60-katanin purified using glutathione-Sepharose
GSTrap column chromatography, GST tag cleaved with PreScission Protease, GST-Sepharose chromatography, gel filtration
-
Ni-NTA column, glutathione-agarose beads
p60, 60 kDa catalytic subunit
-
purification of recombinant protein using a glutathione-sepharose column, removing of the GST moiety with precision protease
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recombinant glutathione S-transferase-tagged form kp60-NTD, affinity purification on glutathione-Sepharose and dialysis
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recombinant His-tagged katanin p60 subunit from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography to over 95% purity
recombinant untagged short isoform of MEI-1 subunit and MBP-TEV-tagged MEI-2 subunit from Escherichia coli by amylose affinity chromatography, eluted with TEV protease
-
using a glutathione Sepharose 4B column
using a glutathione-agarose column, the glutathione-S-transferase moiety is cleaved with thrombin
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using a glutathione-Sepharose column, cleavage of the GST moiety with PreScission Protease
-
using Ni-NTA agarose affinity chromatography
-