Date of Original Version
The pH (Low) Insertion Peptides (pHLIP® peptides) find application in studies of membrane-associated folding, since spontaneous insertion of these peptides is conveniently triggered by varying pH. Here we employed small angle X-ray scattering (SAXS) to investigate WT pHLIP® peptide oligomeric state in solution at high concentrations and monitor changes in liposome structure upon peptide insertion into the bilayer. We established that even at high concentrations (up to 300 μM) WT pHLIP® peptide at pH 8.0 does not form oligomers higher than tetramers (which exhibit concentration-dependent transfer to monomeric state as it was shown previously). This finding has significance for medical applications, when high concentration of the peptide is injected into blood and diluted in blood circulation. The interaction of WT pHLIP® peptide with liposomes does not alter the unilamellar vesicle structure upon peptide adsorption by lipid bilayer at high pH or upon insertion across the bilayer at low pH. At the same time, SAXS data clearly reflect the insertion of the peptide into the membrane at low pH, which opens the possibility to investigate kinetic process of a polypeptide insertion and exit from the membrane in real time by time-resolved SAXS.
Theyencheri Narayanan, Dhammika Weerakkody, Alexander G. Karabadzhak, Michael Anderson, Oleg A. Andreev, and Yana K. Reshetnyak The Journal of Physical Chemistry B 2016 120 (44), 11484-11491
Available at: http://doi.org/10.1021/acs.jpcb.6b06643