Literature summary for 3.13.2.1 extracted from

Brzezinski, K.; Czyrko, J.; Sliwiak, J.; Nalewajko-Sieliwoniuk, E.; Jaskolski, M.; Nocek, B.; Dauter, Z.
S-adenosyl-L-homocysteine hydrolase from a hyperthermophile (Thermotoga maritima) is expressed in Escherichia coli in inactive form - Biochemical and structural studies (2017), Int. J. Biol. Macromol., 104, 584-596 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene sahH, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3)	Thermotoga maritima

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme free or in complex with adenosine, hanging drop vapour diffusion method, mixing of 0.002 ml of 6 mg/ml protein in 100 mM NaCl, and 25 mM Tris-HCl, pH 8.0, with 0.002 ml of reservoir solution containing 20 mM MgCl2 or CaCl2, 0.1 M sodium acetate, pH 4.6 or pH 5.3, and 30% v/v MPD for crystal structure 1 or 2, respectively, with or without addition of adenosine, 18°C, X-ray diffraction structure determination and analysis at 1.75-1.90 A resolution, molecular replacement using the substrate-binding (residues 18-181 and 355-421) and cofactor-binding (182-354) domains of a model of Homo sapiens SAHase (PDB ID 1LI4) as search model	Thermotoga maritima

Crystallization (Comment)

Organism

purified recombinant enzyme free or in complex with adenosine, hanging drop vapour diffusion method, mixing of 0.002 ml of 6 mg/ml protein in 100 mM NaCl, and 25 mM Tris-HCl, pH 8.0, with 0.002 ml of reservoir solution containing 20 mM MgCl2 or CaCl2, 0.1 M sodium acetate, pH 4.6 or pH 5.3, and 30% v/v MPD for crystal structure 1 or 2, respectively, with or without addition of adenosine, 18°C, X-ray diffraction structure determination and analysis at 1.75-1.90 A resolution, molecular replacement using the substrate-binding (residues 18-181 and 355-421) and cofactor-binding (182-354) domains of a model of Homo sapiens SAHase (PDB ID 1LI4) as search model

Thermotoga maritima

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	thermodynamics and kinetics, recombinant enzyme	Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
S-adenosyl-L-homocysteine + H2O	Thermotoga maritima	-	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	Thermotoga maritima DSM 3109	-	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	Thermotoga maritima ATCC 43589	-	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	Thermotoga maritima JCM 10099	-	L-homocysteine + adenosine	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	O51933	-	-
Thermotoga maritima ATCC 43589	O51933	-	-
Thermotoga maritima DSM 3109	O51933	-	-
Thermotoga maritima JCM 10099	O51933	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3) by nickel affinity chromatography, followed by tag cleavage through TEV protease, dialysis, another step of nickel affinity chromatography, ammonium sulfate fractionation, and ultrafiltration, gel filtration, and again ultrafiltration	Thermotoga maritima

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
S-adenosyl-L-homocysteine + H2O	-	Thermotoga maritima	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	-	Thermotoga maritima DSM 3109	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	-	Thermotoga maritima ATCC 43589	L-homocysteine + adenosine	-	?
S-adenosyl-L-homocysteine + H2O	-	Thermotoga maritima JCM 10099	L-homocysteine + adenosine	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 45000, recombinant enzyme, SDS-PAGE	Thermotoga maritima
More	calorimetric evaluation of heat-induced conformational changes at 85°C and oligomeric state	Thermotoga maritima

Synonyms

Synonyms	Comment	Organism
S-adenosyl-L-homocysteine hydrolase	-	Thermotoga maritima
SAHase	-	Thermotoga maritima

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
additional information	-	the enzyme is inactive at room temperature, mechanism of thermal activation, overview. calorimetric evaluation of heat-induced conformational changes (at 85°C) and oligomeric state	Thermotoga maritima

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Thermotoga maritima

General Information

General Information	Comment	Organism
additional information	overall structure of the inactive form of TmSAHase, overview	Thermotoga maritima