Modulation of gramicidin channel conformation and organization by hydrophobic mismatch in saturated phosphatidylcholine bilayers.
The matching of hydrophobic lengths of integral membrane proteins and the surrounding lipid bilayer is an important factor that influences both structure and function of integral membrane proteins. The ion channel gramicidin is known to be uniquely sensitive to membrane properties such as bilayer thickness and membrane mechanical properties. The functionally important carboxy terminal tryptophan residues of gramicidin display conformation-dependent fluorescence which can be used to monitor gramicidin conformations in membranes [S.S. Rawat, D.A. Kelkar, A. Chattopadhyay, Monitoring gramicidin conformations in membranes: a fluorescence approach, Biophys. J. 87 (2004) 831-843]. We have examined the effect of hydrophobic mismatch on the conformation and organization of gramicidin in saturated phosphatidylcholine bilayers of varying thickness utilizing the intrinsic conformation-dependent tryptophan fluorescence. Our results utilizing steady state and time-resolved fluorescence spectroscopic approaches, in combination with circular dichroism spectroscopy, show that gramicidin remains predominantly in the channel conformation and gramicidin tryptophans are at the membrane interfacial region over a range of mismatch conditions. Interestingly, gramicidin conformation shifts toward non-channel conformations in extremely thick gel phase membranes although it is not excluded from the membrane. In addition, experiments utilizing self quenching of tryptophan fluorescence indicate peptide aggregation in thicker gel phase membranes.
Kelkar
Devaki A
DA
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India.
Chattopadhyay
Amitabha
A
D020106:Acrylamide / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D002942:Circular Dichroism; D006096:Gramicidin / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D007473:Ion Channels / Q000737:chemistry*; D008051:Lipid Bilayers / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism*; D020836:Protein Structure, Quaternary; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014364:Tryptophan / Q000378:metabolism
D007473:Ion Channels; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D006096:Gramicidin; D020106:Acrylamide; D014364:Tryptophan