Expression of the pea gene PSMTA in E. coli. Metal-binding properties of the expressed protein.
The pea (Pisum sativum L.) gene PSMTA has an ORF encoding a predicted protein with sequence similarity to class I metallothioneins (MTS). To examine the metal-binding properties of the PSMTA protein it has been expressed in E. coli as a carboxyterminal extension of glutathione-S-transferase (GST). Metal ions were associated with the expressed protein when purified from lysates of E. coli grown in metal supplemented media. The pH of half-dissociation of Zn, Cd and Cu ions from the recombinant fusion protein was determined to be 5.35, 3.95 and 1.45 respectively, compared with equivalent estimates of 4.50, 3.00 and 1.80 for equine renal MT.
Tommey
A M
AM
Department of Biological Sciences, University of Durham, UK.
Shi
J
J
Lindsay
W P
WP
Urwin
P E
PE
Robinson
N J
NJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D001667:Binding, Competitive; D002414:Cations, Monovalent; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D007887:Fabaceae / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005978:Glutathione / Q000737:chemistry; D006859:Hydrogen / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D008969:Molecular Sequence Data; D010946:Plants, Medicinal; D016133:Polymerase Chain Reaction
D002414:Cations, Monovalent; D008670:Metals; D006859:Hydrogen; D008668:Metallothionein; D005978:Glutathione