The binding specificity of OppA determines the selectivity of the oligopeptide ATP-binding cassette transporter.
The purification and functional reconstitution of a five-component oligopeptide ATP-binding cassette transporter with a remarkably wide substrate specificity are described. High-affinity peptide uptake was dependent on liganded substrate-binding protein OppA, which interacts with the translocator OppBCDF with higher affinity than unliganded OppA. Transport screening with combinatorial peptide libraries revealed that (i) the Opp transporter is not selective with respect to amino acid side chains of the transported peptides; (ii) any peptide that can bind to OppA is transported via Opp, including very long peptides up to 35 residues long; and (iii) the binding specificity of OppA largely determines the overall transport selectivity.
Doeven
Mark K
MK
Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, The Netherlands.
Abele
Rupert
R
Tampé
Robert
R
Poolman
Bert
B
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000596:Amino Acids / Q000737:chemistry; D001426:Bacterial Proteins; D001692:Biological Transport; D001920:Bradykinin / Q000493:pharmacokinetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015151:Immunoblotting; D013294:Lactococcus lactis / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry* / Q000378:metabolism; D008715:Methionine / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014633:Valine / Q000737:chemistry
D000596:Amino Acids; D001426:Bacterial Proteins; D002352:Carrier Proteins; D008074:Lipoproteins; D009842:Oligopeptides; D019151:Peptide Library; D010455:Peptides; C046506:oligopeptide-binding protein, bacteria; D000255:Adenosine Triphosphate; D008715:Methionine; D014633:Valine; D001920:Bradykinin