Subconductance states in OmpF gating.
Discrepancies were noted in the published conductance of the Escherichia coli porin OmpF. Results from various papers are hard to compare because of the use of different channel preparations, salt types and concentrations, and electrophysiological techniques (black lipid membrane (BLM) vs. patch clamp). To reconcile these data, we present a side-by-side comparison of OmpF activity studied with the two techniques on the same preparation of pure protein, and in the same low salt concentrations (150 mM KCl). The novel aspect of OmpF porin behavior revealed by this comparison is the ubiquitous existence of states of smaller conductance than the monomeric conductance (subconductance states), regardless of the techniques or experimental conditions used, and the drastic enhancement of subconductance gating by polyamines. Transitions to subconductance states have received little attention in previous publications, in particular when BLM electrophysiology was used. Monomeric closures are rare in recordings at clamped potentials, at least at voltages lower than approximately 100-120 mV. Most closing activity is in the form of subconductance gating, which becomes more dominant in the presence of spermine, with a more frequent and prolonged occupation of these substates. A discussion of the molecular basis for this hallmark behavior of porin is presented.
Baslé
Arnaud
A
Department of Biology and Biochemistry, University of Houston, 369 Science and Research Building 2, Houston, TX 77204-5001, USA.
Iyer
Ram
R
Delcour
Anne H
AH
D002462:Cell Membrane / Q000378:metabolism; D004553:Electric Conductivity; D004594:Electrophysiology; D004926:Escherichia coli / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry; D018408:Patch-Clamp Techniques; D011073:Polyamines / Q000737:chemistry; D018272:Porins / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012492:Salts / Q000494:pharmacology; D013997:Time Factors
D008051:Lipid Bilayers; D008081:Liposomes; C109237:OmpF protein; D011073:Polyamines; D018272:Porins; D012492:Salts