Literature summary for 3.2.1.4 extracted from

Urresti, S.; Cartmell, A.; Liu, F.; Walton, P.H.; Davies, G.J.
Structural studies of the unusual metal-ion site of the GH124 endoglucanase from Ruminiclostridium thermocellum (2018), Acta Crystallogr. Sect. F, 74, 496-505 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS	Acetivibrio thermocellus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged enzyme in complex with G3 or G5f, sitting drop vapour diffuson method, mixing of 0.027 ml of 40 mg/ml protein in 20 mM Tris, pH 7.5, and 6.7 mM G3 or G5f, with 0.027 ml of reservoir solution containing 12% Tacsimate, pH 5.0, 18% or 20% PEG 3350, 2 mM manganese(II) acetate, respectively, and equilibration against 0.4 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.04 and 0.99 A resolution, respectively, modelling	Acetivibrio thermocellus

Crystallization (Comment)

Organism

purified recombinant detagged enzyme in complex with G3 or G5f, sitting drop vapour diffuson method, mixing of 0.027 ml of 40 mg/ml protein in 20 mM Tris, pH 7.5, and 6.7 mM G3 or G5f, with 0.027 ml of reservoir solution containing 12% Tacsimate, pH 5.0, 18% or 20% PEG 3350, 2 mM manganese(II) acetate, respectively, and equilibration against 0.4 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.04 and 0.99 A resolution, respectively, modelling

Acetivibrio thermocellus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	enzyme binding structure, overview	Acetivibrio thermocellus
additional information	the enzyme contains an unusual metal-ion site, which is originally modelled as a Ca2+ site (PDB ID 2xqo) but features aspartic acid, asparagine and two histidine imidazoles as coordinating residues, which are more consistent with a transition-metal binding environment. This metal-ion site can accommodate a range of transition metals, i.e. Fe2+, Cu2+, Mn2+ and Ni2+, but not Ca2+, whilst the three-dimensional structure and mass spectrometry show that one of the histidines is partially covalently modified and is present as a 2-oxohistidine residue, a feature that is rarely observed but that is believed to be involved in an off-switch to transition-metal binding. Atomic resolution complexes define the metal-ion site and also reveal the binding of an unusual fructosylated oligosaccharide, which is presumably present as a contaminant in the cellohexaose used for crystallization	Acetivibrio thermocellus

Organism

Organism	UniProt	Comment	Textmining
Acetivibrio thermocellus	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus ATCC 27405	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus DSM 1237	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus NBRC 103400	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus NCIMB 10682	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus NRRL B-4536	A3DCJ4	i.e. Ruminiclostridium thermocellum	-
Acetivibrio thermocellus VPI 7372	A3DCJ4	i.e. Ruminiclostridium thermocellum	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS, cleavage of the His-tag by 3C protease	Acetivibrio thermocellus

Synonyms

Synonyms	Comment	Organism
Cthe_0435	-	Acetivibrio thermocellus
Dockerin type 1	UniProt	Acetivibrio thermocellus
GH124 endoglucanase	-	Acetivibrio thermocellus
RtGH124	-	Acetivibrio thermocellus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	enzyme melting transition occurs at 72°C in presence of EDTA, Ca2+ does not affect the melting temperature	Acetivibrio thermocellus

General Information

General Information	Comment	Organism
additional information	modelling of the metal-ion sites in the protein three-dimensional structure, overview	Acetivibrio thermocellus