Literature summary for 5.3.3.1 extracted from

Thomas, J.L.; Mason, J.I.; Blanco, G.; Veisaga, M.L.
The engineered, cytosolic form of human type I 3beta-hydroxysteroid dehydrogenase/isomerase: purification, characterization and crystallization (2001), J. Mol. Endocrinol., 27, 77-83.

Cloned(Commentary)

Cloned (Comment)	Organism
insect Sf9 cells transfected with recombinant baculovirus	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging-drop vapor diffusion method. The inability to crystallize the detergent-solubilized, wild-type microsomal enzyme is overcome by engineering a cytosolic form of this protein. Modified enzyme in which the membrane-spanning domain, residues 283-310 of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells	Homo sapiens

Crystallization (Comment)

Organism

hanging-drop vapor diffusion method. The inability to crystallize the detergent-solubilized, wild-type microsomal enzyme is overcome by engineering a cytosolic form of this protein. Modified enzyme in which the membrane-spanning domain, residues 283-310 of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
DELTA283-310	the molecular weight of the subunit is 38800 Da compared to 42000 Da for the wild-type enzyme	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0252	-	5-androstene-3,17-dione	22°C, pH 7.4, truncated cytosolic enzyme DEALTA283-310	Homo sapiens
0.0279	-	5-androstene-3,17-dione	22°C, pH 7.4, wild-type enzyme	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells	Homo sapiens	5829	-
membrane	integral membrane protein	Homo sapiens	16020	-
microsome	-	Homo sapiens	-	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
38800	-	x * 38800, truncated soluble enzyme DEALTA283-310	Homo sapiens
42000	-	x * 42000, wild-type enzyme	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.622	-	reaction with 5-androsten-3,17-dione, truncated cytosolic enzyme DELTA283-310	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5-Androstene-3,17-dione	-	Homo sapiens	4-Androstene-3,17-dione	-	?

Subunits

Subunits	Comment	Organism
?	x * 38800, truncated soluble enzyme DEALTA283-310	Homo sapiens
?	x * 42000, wild-type enzyme	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
NADH	the isomerase activity of the wild-type enzyme and the deletion mutant enzyme DELTA283-310 requires allosteric activation by NADH, Km for wild-type enzyme and deletion mutant enzyme is 0.0046 mM	Homo sapiens