Properties and function of the P type ion pumps cloned from Helicobacter pylori.
Three distinct P type pumps were cloned from H. pylori 69A. Two of these pumps, ATPase 439 and ATPase 948 (CopA), were isolated by gene library screening using DNA oligonucleotide primers. Amino acid similarities found for the predicted proteins were about 50% to Cd2+/Cu2+ pumps. Gene disruption mutagenesis rendered the H. pylori knockout mutants more sensitive to Zn2+ and Cd2+ (ATPase 439) or Cu2+ (CopA). Some of the ATPase 439-deficient mutants were negative for urease activity while the majority of the mutants remained positive. Functional diversity of the pumps was also reflected by the ion affinities found for N-terminal peptides of CopA to Cu2+ and of ATPase 439 to Ni2+, Cu2+ and CO2+. The membrane domain of the two pumps were experimentally shown to consist of eight membrane spans. When ATPase 439 was expressed under control of a tac promoter in Escherichia coli, vanadate-sensitive phosphate accumulation was observed cytochemically along the membrane of the host cells. The third P type pump (ATPase 115) which also exhibited homology to transition metal ATPase was identified by sequencing a library of H. pylori membrane genes. The hydropathy plot of this pump was very similar to the former H. pylori ATPases whereas the N-terminal ion binding region was distinct. It was concluded that, in H. pylori, the presence of three transition metal ATPases with distinct ion specificity contributes to the adaptive mechanisms for gastric survival.
Melchers
K
K
Department of Molecular Biology, Byk Gulden Pharmaceuticals, Konstanz, Germany. melchers@byk.de
Herrmann
L
L
Mauch
F
F
Bayle
D
D
Heuermann
D
D
Weitzenegger
T
T
Schuhmacher
A
A
Sachs
G
G
Haas
R
R
Bode
G
G
Bensch
K
K
Schäfer
K P
KP
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000483:Alleles; D000595:Amino Acid Sequence / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology; D016480:Helicobacter pylori / Q000378:metabolism*; D016623:Ion Pumps / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011487:Protein Conformation
D016623:Ion Pumps; D000251:Adenosine Triphosphatases