Literature summary for 4.4.1.16 extracted from

Collins, R.; Johansson, A.L.; Karlberg, T.; Markova, N.; van den Berg, S.; Olesen, K.; Hammarstroem, M.; Flores, A.; Schueler, H.; Schiavone, L.H.; Brzezinski, P.; Arner, E.S.; Hoegbom, M.
Biochemical discrimination between selenium and sulfur 1: a single residue provides selenium specificity to human selenocysteine lyase (2012), PLoS ONE, 7, e30581.

Cloned(Commentary)

Cloned (Comment)	Organism
sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme in dimeric and tetrameric forms, sitting drop vapour diffusion method, 100 nl of 17 mg/ml protein solution with 100 nl of well solution 1 containing 100 mM HEPES, pH 6.7, and 10% PEG 6000, or well solution 2 containing 50 mM HEPES, pH 8.1, 200 mM ammonium nitrate and 25% PEG 3350, 20°C, 3 days or 7 days, respectively, X-ray diffraction structure determination and analysis of the two crystals forms at 2.1 A and 1.8 A resolution, respectively, modeling of dimeric enzyme complexed with pyridoxal 5'-phosphate	Homo sapiens

Crystallization (Comment)

Organism

purified enzyme in dimeric and tetrameric forms, sitting drop vapour diffusion method, 100 nl of 17 mg/ml protein solution with 100 nl of well solution 1 containing 100 mM HEPES, pH 6.7, and 10% PEG 6000, or well solution 2 containing 50 mM HEPES, pH 8.1, 200 mM ammonium nitrate and 25% PEG 3350, 20°C, 3 days or 7 days, respectively, X-ray diffraction structure determination and analysis of the two crystals forms at 2.1 A and 1.8 A resolution, respectively, modeling of dimeric enzyme complexed with pyridoxal 5'-phosphate

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D146K	site-directed mutagenesis, the mutant variant shows loss of selenocysteine specificity and appearance of cysteine desulfurase activity	Homo sapiens
D146K/H389T	site-directed mutagenesis,the mutant variant shows loss of selenocysteine specificity and appearance of cysteine desulfurase activity to a higher extent	Homo sapiens
H389T	site-directed mutagenesis, the mutant variant shows loss of selenocysteine specificity and appearance of cysteine desulfurase activity	Homo sapiens
V256S	site-directed mutagenesis, the mutant variant shows loss of selenocysteine specificity and appearance of cysteine desulfurase activity	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-selenocysteine + reduced acceptor	Homo sapiens	-	selenide + L-alanine + acceptor	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q96I15	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, followed by tag cleavage through TEV protease, another step of nickel affinity chromatography, and dialysis	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-selenocysteine + reduced acceptor	-	Homo sapiens	selenide + L-alanine + acceptor	-	?
additional information	the enzyme is specific for L-selenocysteine versus L-cysteine, substrate specificity for selenium and sulfur is determined by a single residue, Asp246, in human selenocysteine lyase	Homo sapiens	?	-	?

Synonyms

Synonyms	Comment	Organism
SCL	-	Homo sapiens
Sec lyase	-	Homo sapiens

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
80	-	purified enzyme, pH 8.5, 2 min, inactivation	Homo sapiens

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	-	Homo sapiens

General Information

General Information	Comment	Organism
additional information	the enzyme adopts the canonical fold type I structure of pyridoxal 5'-phosphate enzymes with an active site cysteine, C388	Homo sapiens