Cysteine synthase (CYSM) of Mycobacterium tuberculosis is an O-phosphoserine sulfhydrylase: Evidence for an alternative cysteine biosynthesis pathway in mycobacteria

Agren, D.; Schnell, R.; Oehlmann, W.; Singh, M.; Schneider, G.; J. Biol. Chem. 283, 31567-31574 (2008)

Data extracted from this reference:

Cloned(Commentary)

Commentary	Organism
expressed in Escherichia coli BL21(DE3) cells; expression in Escherichia coli	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization	Organism
2.1 A resolution. A model of O-phosphoserine bound to the enzyme suggests a hydrogen bonding interaction of the side chain of Arg220 with the phosphate group as a key feature in substrate selectivity; sitting drop vapour diffusion method, with 0.1 M Tris-HCl pH 7.25-7.5, 0.1 M K2HPO4, 4.3 M NaCl	Mycobacterium tuberculosis

Engineering

Amino acid exchange	Commentary	Organism
R220A	700fold lower activity with O-phospho-L-serine as substrate compared to the wild type enzyme; significant loss in specificity for substrate O-phosphoserine. The purified R220A mutant shows an absorption spectrum identical to wild type CysM with an absorption band at 412 nm reflecting the Schiff base between Lys51 and PLP. Formation of the aminoacrylate intermediate from O-phospho-L-serine in the mutant is severely compromised, with an approximately 700fold slower rate	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Commentary (Nat. Sub.)	Natural Products	Commentary (Nat. Pro.)	Organism (Nat. Pro.)	Reversibility
More	Mycobacterium tuberculosis	enzyme is involved in an O-phosphoserine based cysteine biosynthesis pathway in Mycobacterium tuberculosis that is independent of both O-acetylserine and the sulphate reduction pathway	?	-	-	-
O-phospho-L-serine + hydrogen sulfide	Mycobacterium tuberculosis	-	L-cysteine + phosphate	-	-	?

Organism

Organism	Primary Accession No. (UniProt)	Commentary	Textmining
Mycobacterium tuberculosis	-	-	-
Mycobacterium tuberculosis	P63873	-	-

Purification (Commentary)

Commentary	Organism
Ni-NTA column chromatography and Superdex-200 gel filtration	Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates	Commentary Substrates	Literature (Substrates)	Organism	Products	Commentary (Products)	Literature (Products)	Organism (Products)	Reversibility
More	enzyme is involved in an O-phosphoserine based cysteine biosynthesis pathway in Mycobacterium tuberculosis that is independent of both O-acetylserine and the sulphate reduction pathway	687773	Mycobacterium tuberculosis	?	-	-	-	-
More	O-acetylserine is not a substrate. Enzyme does not catalyze the reaction of EC 2.5.1.47, O-acetylserine sulfhydrolases	687773	Mycobacterium tuberculosis	?	-	-	-	-
O-acetyl-L-serine + hydrogen sulfide	-	687773	Mycobacterium tuberculosis	L-cysteine + acetate	-	-	-	?
O-phospho-L-serine + hydrogen sulfide	-	687773	Mycobacterium tuberculosis	L-cysteine + phosphate	-	-	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Commentary	Organism	Structure
0.1	-	O-phospho-L-serine	37°C	Mycobacterium tuberculosis

Cofactor

Cofactor	Commentary	Organism	Structure
pyridoxal 5'-phosphate	; bound via a covalent linkage to the side chain of Lys51	Mycobacterium tuberculosis

Cloned(Commentary) (protein specific)

Commentary	Organism
expressed in Escherichia coli BL21(DE3) cells	Mycobacterium tuberculosis
expression in Escherichia coli	Mycobacterium tuberculosis

Cofactor (protein specific)

Cofactor	Commentary	Organism	Structure
pyridoxal 5'-phosphate	bound via a covalent linkage to the side chain of Lys51	Mycobacterium tuberculosis
pyridoxal 5'-phosphate	-	Mycobacterium tuberculosis

Crystallization (Commentary) (protein specific)

Crystallization	Organism
2.1 A resolution. A model of O-phosphoserine bound to the enzyme suggests a hydrogen bonding interaction of the side chain of Arg220 with the phosphate group as a key feature in substrate selectivity	Mycobacterium tuberculosis
sitting drop vapour diffusion method, with 0.1 M Tris-HCl pH 7.25-7.5, 0.1 M K2HPO4, 4.3 M NaCl	Mycobacterium tuberculosis

Engineering (protein specific)

Amino acid exchange	Commentary	Organism
R220A	significant loss in specificity for substrate O-phosphoserine. The purified R220A mutant shows an absorption spectrum identical to wild type CysM with an absorption band at 412 nm reflecting the Schiff base between Lys51 and PLP. Formation of the aminoacrylate intermediate from O-phospho-L-serine in the mutant is severely compromised, with an approximately 700fold slower rate	Mycobacterium tuberculosis
R220A	700fold lower activity with O-phospho-L-serine as substrate compared to the wild type enzyme	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates) (protein specific)

Natural Substrates	Organism	Commentary (Nat. Sub.)	Natural Products	Commentary (Nat. Pro.)	Organism (Nat. Pro.)	Reversibility
More	Mycobacterium tuberculosis	enzyme is involved in an O-phosphoserine based cysteine biosynthesis pathway in Mycobacterium tuberculosis that is independent of both O-acetylserine and the sulphate reduction pathway	?	-	-	-
O-phospho-L-serine + hydrogen sulfide	Mycobacterium tuberculosis	-	L-cysteine + phosphate	-	-	?

Purification (Commentary) (protein specific)

Commentary	Organism
Ni-NTA column chromatography and Superdex-200 gel filtration	Mycobacterium tuberculosis

Substrates and Products (Substrate) (protein specific)

Substrates	Commentary Substrates	Literature (Substrates)	Organism	Products	Commentary (Products)	Literature (Products)	Organism (Products)	Reversibility
More	enzyme is involved in an O-phosphoserine based cysteine biosynthesis pathway in Mycobacterium tuberculosis that is independent of both O-acetylserine and the sulphate reduction pathway	687773	Mycobacterium tuberculosis	?	-	-	-	-
More	O-acetylserine is not a substrate. Enzyme does not catalyze the reaction of EC 2.5.1.47, O-acetylserine sulfhydrolases	687773	Mycobacterium tuberculosis	?	-	-	-	-
O-acetyl-L-serine + hydrogen sulfide	-	687773	Mycobacterium tuberculosis	L-cysteine + acetate	-	-	-	?
O-phospho-L-serine + hydrogen sulfide	-	687773	Mycobacterium tuberculosis	L-cysteine + phosphate	-	-	-	?
O-phospho-L-serine + hydrogen sulfide	-	687773	Mycobacterium tuberculosis	L-cysteine + phosphate	-	-	-	?

Turnover Number [1/s] (protein specific)

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Commentary	Organism	Structure
0.1	-	O-phospho-L-serine	37°C	Mycobacterium tuberculosis

See also following references to EC number 2.5.1.65 (sorted by year of publication):

No.	1st author	Pub Med	title	organims	journal	volume	pages	year	Activating Compound	Application	Cloned(Commentary)	Crystallization (Commentary)	Engineering	General Stability	Inhibitors	KM Value [mM]	Localization	Metals/Ions	Molecular Weight [Da]	Natural Substrates/ Products (Substrates)	Organic Solvent Stability	Organism	Oxidation Stability	Posttranslational Modification	Purification (Commentary)	Reaction	Renatured (Commentary)	Source Tissue	Specific Activity [micromol/min/mg]	Storage Stability	Substrates and Products (Substrate)	Subunits	Temperature Optimum [°C]	Temperature Range [°C]	Temperature Stability [°C]	Turnover Number [1/s]	pH Optimum	pH Range	pH Stability	Cofactor	Ki Value [mM]	pI Value	IC50 Value	Activating Compound (protein specific)	Application (protein specific)	Cloned(Commentary) (protein specific)	Cofactor (protein specific)	Crystallization (Commentary) (protein specific)	Engineering (protein specific)	General Stability (protein specific)	IC50 Value (protein specific)	Inhibitors (protein specific)	Ki Value [mM] (protein specific)	KM Value [mM] (protein specific)	Localization (protein specific)	Metals/Ions (protein specific)	Molecular Weight [Da] (protein specific)	Natural Substrates/ Products (Substrates) (protein specific)	Organic Solvent Stability (protein specific)	Oxidation Stability (protein specific)	Posttranslational Modification (protein specific)	Purification (Commentary) (protein specific)	Renatured (Commentary) (protein specific)	Source Tissue (protein specific)	Specific Activity [micromol/min/mg] (protein specific)	Storage Stability (protein specific)	Substrates and Products (Substrate) (protein specific)	Subunits (protein specific)	Temperature Optimum [°C] (protein specific)	Temperature Range [°C] (protein specific)	Temperature Stability [°C] (protein specific)	Turnover Number [1/s] (protein specific)	pH Optimum (protein specific)	pH Range (protein specific)	pH Stability (protein specific)	pI Value (protein specific)	Expression	General Information	General Information (protein specific)	Expression (protein specific)	KCat/KM [mM/s]	KCat/KM [mM/s] (protein specific)
685195	OLeary		O-Phospho-l-serine and the thi ...	Mycobacterium tuberculosis	Biochemistry	47	11606-11615	2008	-	-	-	-	-	-	-	-	-	-	-	-	-	1	-	-	-	-	-	-	-	-	3	-	-	-	-	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	3	-	-	-	-	2	-	-	-	-	-	-	-	-	-	-
687773	Agren		Cysteine synthase (CYSM) of My ...	Mycobacterium tuberculosis	J. Biol. Chem.	283	31567-31574	2008	-	-	1	1	1	-	-	-	-	-	-	2	-	4	-	-	1	-	-	-	-	-	4	-	-	-	-	1	-	-	-	1	-	-	-	-	-	2	2	2	2	-	-	-	-	-	-	-	-	2	-	-	-	1	-	-	-	-	5	-	-	-	-	1	-	-	-	-	-	-	-	-	-	-
659728	Oda		Three-dimensional structure of ...	Aeropyrum pernix	J. Mol. Biol.	351	334-344	2005	-	-	1	1	1	-	-	-	-	-	-	1	-	4	-	-	1	1	-	-	4	-	4	1	1	-	-	-	-	-	-	1	-	-	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	1	-	-	-	1	-	-	4	-	4	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-
637385	Mino		Characterization of a novel th ...	Aeropyrum pernix	J. Bacteriol.	185	2277-2284	2003	-	-	1	-	-	-	-	3	-	-	1	1	-	1	-	-	1	-	-	-	1	1	4	1	-	-	1	2	1	-	4	1	-	-	-	-	-	1	1	-	-	-	-	-	-	3	-	-	1	1	-	-	-	1	-	-	1	1	4	1	-	-	1	2	1	-	4	-	-	-	-	-	-	-
654085	Mino		Crystallization and preliminar ...	Aeropyrum pernix	Acta Crystallogr. Sect. D	59	338-340	2003	-	-	1	1	-	-	-	-	-	-	-	3	-	1	-	-	1	1	-	-	-	-	4	1	-	-	-	-	-	-	-	1	-	-	-	-	-	1	1	1	-	-	-	-	-	-	-	-	-	3	-	-	-	1	-	-	-	-	4	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
655576	Mino		A novel O-phospho-L-serine sul ...	Aeropyrum pernix	FEBS Lett.	551	133-138	2003	1	-	1	-	-	-	23	10	-	-	-	3	-	1	-	-	1	1	-	-	1	-	13	1	3	-	1	3	1	-	-	1	-	-	-	1	-	1	1	-	-	-	-	23	-	10	-	-	-	3	-	-	-	1	-	-	1	-	13	1	3	-	1	3	1	-	-	-	-	-	-	-	-	-