Purification and characterization of a glycine betaine binding protein from Escherichia coli.
A major component of the Escherichia coli response to elevated medium osmolarity is the synthesis of a periplasmic protein with an Mr of 31,000. The protein was absent in mutants with lambda placMu insertions in the proU region, a locus involved in transport of the osmoprotectant glycine betaine. This periplasmic protein has now been purified to homogeneity. Antibody directed against the purified periplasmic protein crossreacts with the fusion protein produced as a result of the lambda placMu insertion, indicating that proU is the structural gene specifying the 31-kDa protein. The purified protein binds glycine betaine with high affinity but has no affinity for either proline or choline, clarifying the role of proU in osmoprotectant transport. The amino-terminal sequence of the mature glycine betaine binding protein is Ala-Asp-Leu-Pro-Gly-Lys-Gly-Ile-Thr-Val-Asn-Pro.
Barron
A
A
Jung
J U
JU
Villarejo
M
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D001622:Betaine / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000032:analysis* / Q000235:genetics; D029968:Escherichia coli Proteins; D005796:Genes; D005798:Genes, Bacterial; D026901:Membrane Transport Proteins; D008970:Molecular Weight; D009994:Osmolar Concentration; D033902:Periplasmic Binding Proteins; D010957:Plasmids; D011392:Proline / Q000378:metabolism; D011401:Promoter Regions, Genetic; D013347:Subcellular Fractions / Q000032:analysis
D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C053437:PROX protein, E coli; D033902:Periplasmic Binding Proteins; D001622:Betaine; D011392:Proline