Synthetic polypeptide adsorption to Cu-IDA containing lipid films: a model for protein-membrane interactions.
Adsorption of synthetic alanine-rich peptides to lipid monolayers was studied by X-ray and neutron reflectivity, grazing incidence X-ray diffraction (GIXD), and circular dichroic spectroscopy. The peptides contained histidine residues to drive adsorption to Langmuir monolayers of lipids with iminodiacetate headgroups loaded with Cu2+. Adsorption was found to be irreversible with respect to bulk peptide concentration. The peptides were partially helical in solution at room temperature, the temperature of the adsorption assays. Comparisons of the rate of binding and the structure of the adsorbed layer were made as a function of the number of histidines (from 0 to 2) and also as a function of the positioning of the histidines along the backbone. For peptides containing two histidines on the same side of the helical backbone, large differences were observed in the structure of the adsorbed layer as a function of the spacing of the histidines. With a spacing of 6 A, there was a substantial increase in helicity upon binding (from 17% to 31%), and the peptides adsorbed to a final density approaching that of a nearly completed monolayer of alpha-helices adsorbed side-on. The thickness of the adsorbed layer (17 +/- 2.5 A) was slightly greater than the diameter of alpha-helices, suggesting that the free, unstructured ends extended into solution. With a spacing of 30 A between histidines, a far weaker increase in helicity upon binding was observed (from 13% to 19%) and a much lower packing density resulted. The thickness of the adsorbed layer (10 +/- 4 A) was smaller, consistent with the ends being bound to the monolayer. Striking differences were observed in the interaction of the two types of peptide with the lipid membrane by GIXD, consistent with binding by two correlated sites only for the case of 6 A spacing. All these results are attributed to differences in spatial correlation between the histidines as a function of separation distance along the backbone for these partially helical peptides. Finally, control over orientation was demonstrated by placing a histidine on an end of the sequence, which resulted in adsorbed peptides oriented perpendicular to the membrane.
Kent
M S
MS
Sandia National Laboratories, Albuquerque, New Mexico, USA.
Yim
H
H
Murton
J K
JK
Sasaki
D Y
DY
Polizzotti
B D
BD
Charati
M B
MB
Kiick
K L
KL
Kuzmenko
I
I
Satija
S
S
D000327:Adsorption; D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003300:Copper; D015195:Drug Design; D006639:Histidine / Q000737:chemistry; D007098:Imino Acids; D008081:Liposomes / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D013057:Spectrum Analysis; D014965:X-Rays
D007098:Imino Acids; D008081:Liposomes; D008563:Membrane Lipids; D008565:Membrane Proteins; D010455:Peptides; D006639:Histidine; D003300:Copper; C008109:iminodiacetic acid