Argüello, José M.
The goal of our studies was the cloning and characterization of heavy metal transporting P-type ATPases. This subfamily of P-type ATPases is responsible for metal biotolerance and bioaccumulation. Two different heavy metal transporting ATPases were the target of our studies - a putative Cu? ATPase encoded by the open reading frame HI0290 in Haemophilus influenzae and a putative Cu? ATPase (pacS) found in Archaeoglobus fulgidus. Polymerase chain reaction (PCR) methods were used to amplify the cDNAs encoding these enzymes. The cDNAs were cloned into a pGEM vector, and then subcloned into bacterial expression vector pET-21b. In addition, alignment studies, done with the amino acid sequences of currently known heavy metal transporting P-type ATPases, reveal six major groupswithin this subfamily with distinct differences. The differences observed in transmembrane helices H7 and H8 appear to be the basis of ion selectivity by these enzymes. Current studies are focused on the expression and functional characterization of these proteins.
Worcester Polytechnic Institute
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Chemistry and Biochemistry