Molecular Cloning, Heterologous Expression, and Characterization of Human Glyoxalase II

Purification, molecular cloning, heterologous expression and characterization of pig CYP1A2.

Porcine cytochrome P450 (CYP) 1A2 was purified to electrophoretic homogeneity from the hepatic microsomes of beta-naphthoflavone-treated male pigs. In a reconstituted system, this enzyme showed a good catalytic activity towards caffeine, acetanilide, and methoxyresorufin, all known markers of mammalian CYP1A2. Using 3'- and 5'-rapid amplification of coding DNA (cDNA) ends (RACE), we amplified f...

Cloning, heterologous expression, and enzymological characterization of human squalene monooxygenase.

The cDNA for human squalene monooxygenase, a key enzyme in the committed pathway for cholesterol biosynthesis, was amplified from a human liver cDNA library and cloned, and the protein was expressed in Escherichia coli and purified. Kinetic analysis of the purified enzyme revealed an apparent K(m) for squalene of 7.7 microM and an apparent k(cat) of 1.1 min(-1). For FAD the apparent K(m) is 0.3...

Molecular cloning and characterization of the thiolesterase glyoxalase II from Arabidopsis thaliana.

cDNA encoding glyoxalase II from Arabidopsis thaliana has been cloned and sequenced. The isolated 894 bp segment included a sequence of 774 bp encoding a protein with a calculated molecular mass of 28,791 Da. The amino acid sequence deduced from the A. thaliana cDNA showed 54% identity with that of the human enzyme. Searches in databanks identified seven additional DNA sequences from different ...

Cloning and characterization of human colon glyoxalase-I.

Glyoxalase-I cDNA clones were isolated from a human colon cDNA library using polyclonal antibodies raised against the protein purified from human colon tissue. Positive clones were purified, subcloned, and their nucleotide sequence determined. The glyoxalase-I cDNA encodes a 184-amino acid protein with a predicted molecular weight of 20,774, corresponding to the monomeric subunit weight of the ...

Homology Cloning, Heterologous Expression and Characterization of a New Channelrhodopsin