Literature summary for 3.6.4.B10 extracted from

Shah, R.; Large, A.T.; Ursinus, A.; Lin, B.; Gowrinathan, P.; Martin, J.; Lund, P.A.
Replacement of GroEL in Escherichia coli by the group II chaperonin from the archaeon Methanococcus maripaludis (2016), J. Bacteriol., 198, 2692-2700 .

Search for more literature for 3.6.4.B10

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of archaeal group II chaperonin Mm-cpn in Escherichia coli strain TAB21 lacking its group I chaperonin GroEL, and wild-type and mutant enzymes overexpression in Escherichia coli strain BL21(DE3)	Methanococcus maripaludis

Protein Variants

Protein Variants	Comment	Organism
D386A	site-directed mutagenesis, introduction of D386A into Mm-cpn significantly reduces its ability to complement for loss of GroES and GroEL, loss of ATPase activity severely affects the complementing ability of the wild-type and mutant Mm-cpn proteins	Methanococcus maripaludis
K216A	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216C	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216D	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216E	random mutagenesis, growth for the mutants is clearly faster than for wild-type Mm-cpn organisms under GroES/GroEL-limiting conditions, improved phenotype in Escherichia coli under GroEL- and GroES-depleting conditions. The mutant can effectively hydrolyze ATP	Methanococcus maripaludis
K216E/D386A	site-directed mutagenesis, introduction of D386A into Mm-cpn significantly reduces its ability to complement for loss of GroES and GroEL, loss of ATPase activity severely affects the complementing ability of the wild-type and mutant Mm-cpn proteins	Methanococcus maripaludis
K216F	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216G	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216L	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216P	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216Q	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216R	site-directed mutagenesis, the mutant enzyme slightly complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216S	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216T	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
K216V	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
K216Y	site-directed mutagenesis, the mutant enzyme moderately complements the GroEL-deletion mutant Escherichia coli strain TAB21	Methanococcus maripaludis
M223E	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223F	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223G	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223I	random mutagenesis, growth for the mutants is clearly faster than for wild-type Mm-cpn organisms under GroES/GroEL-limiting conditions, improved phenotype in Escherichia coli under GroEL- and GroES-depleting conditions. The mutant can effectively hydrolyze ATP	Methanococcus maripaludis
M223I	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223I/D386A	site-directed mutagenesis, introduction of D386A into Mm-cpn significantly reduces its ability to complement for loss of GroES and GroEL, loss of ATPase activity severely affects the complementing ability of the wild-type and mutant Mm-cpn proteins	Methanococcus maripaludis
M223L	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223R	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223S	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223V	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223W	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
M223Y	site-directed mutagenesis, the mutant enzyme complements the GroEL-deletion mutant Escherichia coli strain TAB21 well	Methanococcus maripaludis
additional information	summary of the functional growth analysis of Escherichia coli TAB21 cells expressing diverse Mm-cpn-M223 and Mm-cpn-K216 mutants at 30°C, overview. The Mm-cpn-K216E and Mm-cpn-M223I mutants act as genuine chaperonins and must complete an ATP-dependent chaperonin cycle to function in Escherichia coli	Methanococcus maripaludis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Methanococcus maripaludis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
900000	-	about, Mm-cpn protein complex, gel filtration and native PAGE	Methanococcus maripaludis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + H2O	Methanococcus maripaludis	-	ADP + phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanococcus maripaludis	Q877G8	i.e. Methanococcus deltae	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant chaperonin Mm-cpn from Escherichia coli strain BL21(DE3) by anion exchange chromatography, ammonium sulfate fractionation, and gel filtation, followed by ultrafiltration	Methanococcus maripaludis

Source Tissue

Source Tissue	Comment	Organism	Textmining
additional information	the optimum growth temperature of the parent organism is 35-40°C	Methanococcus maripaludis	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + H2O	-	Methanococcus maripaludis	ADP + phosphate	-	?

Subunits

Subunits	Comment	Organism
More	Mm-cpn is homooligomeric	Methanococcus maripaludis

Synonyms

Synonyms	Comment	Organism
archaeal chaperonin	-	Methanococcus maripaludis
group II chaperonin	-	Methanococcus maripaludis
Mm-cpn	-	Methanococcus maripaludis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	ATPase assay at	Methanococcus maripaludis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	ATPase assay at	Methanococcus maripaludis

General Information

General Information	Comment	Organism
malfunction	the wild-type organism shows reduced growth under GroES/GroEL-limiting conditions. Cells are transformed with plasmids expressing Mm-cpn, Mm-cpn-M223I, and Mm-cpn-K216E and grown at 26°C, 30°C, and 37°C in the presence of sucrose. Apart from the positive control, growth is observed only for cells expressing the mutant Mm-cpn-K216E. These cells show distinct visible colonies after 3 days at 26°C and 30°C in numbers comparable to those observed for cells expressing GroEL. Cells expressing Mm-cpn-K216E also form visible colonies at 37°C after 5 days. Loss of ATPase activity by mutation D386A severely affects the complementing ability of the wild-type and mutant Mm-cpn proteins	Methanococcus maripaludis
additional information	ATP hydrolysis is important to attain the closed conformation of the enzyme	Methanococcus maripaludis