Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa

Elif Eren, Jamie Parkin, Ayodele Adelanwa, Belete Cheneke, Liviu Movileanu, Syma Khalid, Bert Van Den Berg

Research output: Research - peer-reviewArticle

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Abstract

Because small molecules enter Gram-negative bacteria via outer membrane (OM) channels, understanding OM transport is essential for the rational design of improved and new antibiotics. In the human pathogen Pseudomonas aeruginosa, most small molecules are taken up by outer membrane carboxylate channel (Occ) proteins, which can be divided into two distinct subfamilies, OccD and OccK. Here we characterize substrate transport mediated by Occ proteins belonging to both subfamilies. Based on the determination of the OccK2-glucuronate cocrystal structure, we identify the channel residues that are essential for substrate transport. We further show that the pore regions of the channels are rigid in the OccK subfamily and highly dynamic in the OccD subfamily. We also demonstrate that the substrate carboxylate group interacts with central residues of the basic ladder, a row of arginine and lysine residues that leads to and away from the binding site at the channel constriction. Moreover, the importance of the basic ladder residues corresponds to their degree of conservation. Finally, we apply the generated insights by converting the archetype of the entire family, OccD1, from a basic amino acid-specific channel into a channel with a preference for negatively charged amino acids.

LanguageEnglish (US)
Pages12042-12053
Number of pages12
JournalJournal of Biological Chemistry
Volume288
Issue number17
DOIs
StatePublished - Apr 26 2013

Fingerprint

Membranes
Molecules
Substrates
Ion Channels
Pseudomonas aeruginosa
Ladders
Proteins
Basic Amino Acids
Glucuronic Acid
Pathogens
Lysine
Arginine
Conservation
Bacteria
Binding Sites
Anti-Bacterial Agents
Amino Acids
Gram-Negative Bacteria
Constriction

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Eren, E., Parkin, J., Adelanwa, A., Cheneke, B., Movileanu, L., Khalid, S., & Van Den Berg, B. (2013). Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa. Journal of Biological Chemistry, 288(17), 12042-12053. DOI: 10.1074/jbc.M113.463570

Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa. / Eren, Elif; Parkin, Jamie; Adelanwa, Ayodele; Cheneke, Belete; Movileanu, Liviu; Khalid, Syma; Van Den Berg, Bert.

In: Journal of Biological Chemistry, Vol. 288, No. 17, 26.04.2013, p. 12042-12053.

Research output: Research - peer-reviewArticle

Eren, E, Parkin, J, Adelanwa, A, Cheneke, B, Movileanu, L, Khalid, S & Van Den Berg, B 2013, 'Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa' Journal of Biological Chemistry, vol 288, no. 17, pp. 12042-12053. DOI: 10.1074/jbc.M113.463570
Eren E, Parkin J, Adelanwa A, Cheneke B, Movileanu L, Khalid S et al. Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa. Journal of Biological Chemistry. 2013 Apr 26;288(17):12042-12053. Available from, DOI: 10.1074/jbc.M113.463570
Eren, Elif ; Parkin, Jamie ; Adelanwa, Ayodele ; Cheneke, Belete ; Movileanu, Liviu ; Khalid, Syma ; Van Den Berg, Bert. / Toward understanding the outer membrane uptake of small molecules by Pseudomonas aeruginosa. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 17. pp. 12042-12053
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