PMID: 2496114May 1, 1989Paper

Deletion mutants of the Escherichia coli K-12 mannitol permease: dissection of transport-phosphorylation, phospho-exchange, and mannitol-binding activities

Journal of Bacteriology
P L GrisafiJ Lengeler

Abstract

We have constructed a series of deletion mutations of the cloned Escherichia coli K-12 mtlA gene, which encodes the mannitol-specific enzyme II of the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system. This membrane-bound permease consists of 637 amino acid residues and is responsible for the concomitant transport and phosphorylation of D-mannitol in E. coli. Deletions into the 3' end of mtlA were constructed by exonuclease III digestion. Restriction mapping of the resultant plasmids identified several classes of deletions that lacked approximately 5% to more than 75% of the gene. Immunoblotting experiments revealed that many of these plasmids expressed proteins within the size range predicted by the restriction analyses, and all of these proteins were membrane localized, which demonstrated that none of the C-terminal half of the permease is required for membrane insertion. Functional analyses of the deletion proteins, expressed in an E. coli strain deleted for the chromosomal copy of mtlA, showed that all but one of the strains containing confirmed deletions were inactive in transport and PEP-dependent phosphorylation of mannitol, but deletions removing up to at least 117 amino acid residues from the C...Continue Reading

References

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Citations

Jul 11, 2006·Protein Science : a Publication of the Protein Society·Gertjan VeldhuisBert Poolman
Jan 30, 1990·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·J W LengelerB M Wöhrl
Aug 22, 2003·Protein Science : a Publication of the Protein Society·Jaap BroosGiovanni B Strambini
Dec 13, 2006·Microbiology and Molecular Biology Reviews : MMBR·Josef DeutscherPieter W Postma
Nov 5, 1992·Journal of Molecular Biology·L A LammersG T Robillard
Aug 1, 1992·Protein Expression and Purification·P T Beernink, D R Tolan
Sep 1, 1993·Microbiological Reviews·P W PostmaG R Jacobson

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