Functional analyses reveal an important role for tyrosine residues in the staphylococcal multidrug efflux protein QacA.
BACKGROUND
The staphylococcal QacA multidrug efflux protein confers resistance to an exceptional number of structurally unrelated antimicrobial compounds. Aromatic amino acid residues have been shown to be highly important for the transport function of several multidrug transporters and are intimately involved in multidrug binding. This study investigated the structural and functional importance of the seven tyrosine residues in QacA by examining the phenotypic effect of incorporating conservative (aromatic) and non-conservative (non-aromatic) substitutions for these residues.


RESULTS
Determination of the resistance profiles and analysis of drug transport assays revealed that non-conservative substitutions for most tyrosine residues influenced the QacA drug recognition spectrum. However, an aromatic residue at three tyrosine positions, 63, 410 and 429, was of importance for QacA-mediated transport and resistance to the majority of substrates tested.


CONCLUSIONS
A tyrosine or phenylalanine residue at amino acid positions corresponding to 63 of QacA in related drug efflux proteins is found to be highly conserved. Therefore, an aromatic side chain at this position is likely to partake in a function common to these drug transporters, such as proton translocation or essential intramolecular contacts, whereas aromatic residues at the non-conserved 410 and 429 positions are expected to mediate a QacA-specific function, possibly forming or stabilising part of the QacA drug binding region.
Wu
Jingqin
J
School of Biological Sciences, The University of Sydney, Sydney, New South Wales, 2006, Australia. wujingqin@hotmail.com
Hassan
Karl A
KA
Skurray
Ronald A
RA
Brown
Melissa H
MH
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D004996:Ethidium / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D009154:Mutation / Q000235:genetics; D013210:Staphylococcus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; C068498:qacA protein, Staphylococcus aureus; D014443:Tyrosine; D004996:Ethidium
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