Summary
Commercially obtained cystine binding protein (CBP), an osmotic shock protein ofEscherichia coli, was studied in an effort to determine its binding characteristics. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) analysis of commercially obtained CBP showed three protein bands. N-terminal amino acid microsequencing and subsequent computer search revealed that the sequence of one of these proteins (25-kDa) was nearly identical to histidine binding protein (HisJ) ofSalmonella typhimurium. Purification of CBP by HPLC yielded four protein peaks, of which one bound histidine exclusively. Binding was maximal at pH 5.0 to 6.0, at 4°C, did not require calcium or magnesium ions and was not inhibited by reduction of CBP disulfide bonds. Amino acids other than histidine or cystine did not bind to CBP. These data show that commercially available CBP is not a homogenous protein; it contains a histidine as well as a cystine binding component.
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References
Adams MD, Oxender DL (1989) Bacterial periplasmic binding protein tertiary structures. J Biol Chem 264: 15739–15742
Berger EA, Heppel LA (1972) A binding protein involved in the transport of cystine and diaminopimelic acid inEscherichia coli. J Biol Chem 247: 7684–7694
Higgins CF, Ames GF (1981) Two periplasmic transport proteins which interact with a common membrane receptor show extensive homology: complete nucleotide sequences. Proc Natl Acad Sci USA 78: 6038–6042
Higgins CF, Hyde SC, Mimmack MM, Gileadi U, Fell DR, Gallagher MP (1990) Binding protein-dependent transport systems. J Bioenerget Biomembr 22: 571–591
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage. Nature (London) 227: 680–685
Lipman DJ, Pearson WR (1985) Rapid and sensitive protein similarity searches. Science 227: 1435–1441
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Oshima RG, Willis RC, Furlong CE, Schneider JA (1974) Binding assays for amino acids. The utilization of a cystine binding protein fromEscherichia coli for the determination of acid-soluble cystine in small physiological samples. J Biol Chem 249: 6033–6039
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Butler, J.D., Levin, S.W., Facchiano, A. et al. A histidine binding protein ofEscherichia coli: a component of cystine binding protein ofEscherichia coli . Amino Acids 5, 39–50 (1993). https://doi.org/10.1007/BF00806191
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DOI: https://doi.org/10.1007/BF00806191