PMID: 8611597Jan 12, 1996Paper

Physiological evidence for an interaction between Glu-325 and His-322 in the lactose carrier of Escherichia coli

Biochimica Et Biophysica Acta
J I LeeT H Wilson

Abstract

Site-directed mutagenesis and second-site suppressor analysis have proven to be useful approaches to examine the role of charged amino acids in the structure and function of the lactose carrier of Escherichia coli. A lactose carrier mutant Glu-325 --> Ser failed to ferment melibiose and showed white clones on melibiose MacConkey indicator plates. Several red revertants were isolated from these plates. Two of these revertants showed a double mutation, the original mutation (Glu-325 --> Ser) plus His-322 --> Asp. Seven revertants showed a second site mutation His-322 --> Asn. Although the second site revertants failed to accumulate sugars they do show more rapid uptake of melibiose into cells containing alpha-galactosidase than the original mutant Glu-325 --> Ser. The complete loss of transport activity due to the removal of the negative charge at 325 can be partially compensated for by the introduction of a new negative charge at 322. A site-directed double mutant His-322 --> Asn/Glu-325 --> Asn showed a greater rate of lactose uptake (Vmax) than either of the single mutants His-322 --> Asn or Glu 325 --> Asn. It was concluded that there is some type of physiological interaction (possibly a salt bridge) between His-322 and Glu-325.

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Citations

Jul 1, 1997·Protein Science : a Publication of the Protein Society·J SunH R Kaback
Mar 19, 2009·The Journal of Membrane Biology·Yang PengManuel F Varela
Aug 1, 1997·Current Opinion in Structural Biology·H R KabackJ Wu
Dec 4, 2004·Proceedings of the National Academy of Sciences of the United States of America·Shiela E UnklesJames R Kinghorn
Sep 17, 1996·Proceedings of the National Academy of Sciences of the United States of America·J WuH R Kaback
Dec 10, 1996·Proceedings of the National Academy of Sciences of the United States of America·J Wu, H R Kaback
May 27, 1997·Proceedings of the National Academy of Sciences of the United States of America·H R Kaback
Feb 28, 2007·Biophysical Journal·Richard J NaftalinPhilip Cunningham
Jun 25, 2004·The Journal of Membrane Biology·J L Johnson, R J Brooker
Aug 7, 1996·Biochimica Et Biophysica Acta·M F Varela, T H Wilson

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