Literature summary for 3.1.13.4 extracted from

Wang, H.; Morita, M.; Yang, X.; Suzuki, T.; Yang, W.; Wang, J.; Ito, K.; Wang, Q.; Zhao, C.; Bartlam, M.; Yamamoto, T.; Rao, Z.
Crystal structure of the human CNOT6L nuclease domain reveals strict poly(A) substrate specificity (2010), EMBO J., 29, 2566-2576.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutants of the truncated catalytic domain of human CNOT6L, residues 158-555, in Escherichia coli	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant CNOT6L nuclease domain in complex with AMP and poly(A) DNA, hanging drop vapour diffusion method, 0.001 ml of protein solution, containing 15 mg/ml protein and 1 mM DTT, is mixed with 0.001 ml of reservoir solution and equilibrated over 300 ml reservoir solution, containing 0.1 M HEPES, pH 7.5, 1.1 M ammonium tartrate, and 0.2 M NDSB-201, at 16°C, X-ray diffraction structure determination and analysis at 1.94-2.44 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant CNOT6L nuclease domain in complex with AMP and poly(A) DNA, hanging drop vapour diffusion method, 0.001 ml of protein solution, containing 15 mg/ml protein and 1 mM DTT, is mixed with 0.001 ml of reservoir solution and equilibrated over 300 ml reservoir solution, containing 0.1 M HEPES, pH 7.5, 1.1 M ammonium tartrate, and 0.2 M NDSB-201, at 16°C, X-ray diffraction structure determination and analysis at 1.94-2.44 A resolution

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D489A	site-directed mutagenesis, active site mutant, inactive mutant	Homo sapiens
E240A	site-directed mutagenesis, active site mutant, inactive mutant	Homo sapiens
F484A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
H529A	site-directed mutagenesis, active site mutant, inactive mutant	Homo sapiens
L197A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
L216A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
L414A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
N412A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
P365A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
P365A/F484A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens
P365A/N412A/F484A	site-directed mutagenesis, active site mutant, not expressable in Escherichia coli	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
nucleus	-	Homo sapiens	5634	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	the deadenylase activity of CNOT6L is strictly dependent on the presence of Mg2+	Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
45000	-	x * 45000, recombinant CNOT6L nuclease domain, SDS-PAGE	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Homo sapiens	the nuclease domain of CNOT6L exhibits full Mg2+-dependent deadenylase activity with strict poly(A) RNA substrate specificity. The active site of CNOT6L recognizes the RNA substrate through its negatively charged surface	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant E240A, D489A, and H529A CNOT6L nuclease domains from Escherichia coli	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
exonucleolytic cleavage of poly(A) to 5'-AMP	catalytic mechanism for CNOT6L 3'-5' deadenylation involving a pentacovalent phosphate transition, overview	Homo sapiens	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the nuclease domain of CNOT6L exhibits full Mg2+-dependent deadenylase activity with strict poly(A) RNA substrate specificity. The active site of CNOT6L recognizes the RNA substrate through its negatively charged surface	Homo sapiens	?	-	?
additional information	full-length CNOT6L shows 3'-5' deadenylase activity, the enzyme exhibits a molecular deadenylase mechanism involving a pentacovalent phosphate transition. The 3'-5' deadenylase activity of the CNOT6L catalytic domain alone for poly(A) RNA substrates appears to be increased with 7 nucleotides at the 5' end, or reduced with 25 nucleotides at the 5' end, substrate specificity, overview	Homo sapiens	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 45000, recombinant CNOT6L nuclease domain, SDS-PAGE	Homo sapiens

Synonyms

Synonyms	Comment	Organism
CNOT6L nuclease	-	Homo sapiens
More	the enzyme belongs to the EEP family adopts a complete alpha/beta sandwich fold typical of hydrolases with highly conserved active site residues similar to APE1	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Homo sapiens