PMID:2007584
| Citation |
Forchhammer, K, Leinfelder, W, Boesmiller, K, Veprek, B and Böck, A (1991) Selenocysteine synthase from Escherichia coli. Nucleotide sequence of the gene (selA) and purification of the protein. J. Biol. Chem. 266:6318-23 |
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| Abstract |
The nucleotide sequence of the selA gene from Escherichia coli whose product is involved in the conversion of seryl-tRNA(Sec UCA) into selenocysteyl-tRNA(Sec UCA) was determined. selA codes for a polypeptide of a calculated Mr of 50,667; a protein of appropriate size was synthesized in vivo in a T7 promoter/polymerase system. An assay for SELA activity was devised which is based on the seryl-tRNA(Sec UCA)-dependent incorporation of [75Se] selenium into acid-insoluble material. It was used to follow SELA purification from cells that overproduced the protein from a phage T7 promoter plasmid. Purified native SELA protein migrates in gel filtration experiments with a native Mr of about 600,000. SELA contains 1 mol of bound pyridoxal 5-phosphate/mol of 50-kDa subunit. Evidence is presented that the overall conversion of seryl-tRNA(Sec UCA) to selenocysteyl-tRNA(Sec UCA) occurs at the SELA protein. SELA, therefore, has the function of a selenocysteine synthase. |
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| Keywords |
Amino Acid Sequence; Base Sequence; Chromatography, Liquid; Circular Dichroism; DNA, Bacterial/genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli/enzymology; Molecular Sequence Data; Molecular Weight; Plasmids; Promoter Regions, Genetic; Pyridoxal Phosphate/analysis; RNA, Transfer/genetics; Restriction Mapping; Spectrophotometry, Ultraviolet; Transferases/genetics; Transferases/isolation & purification; Transferases/metabolism |
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