Photo-crosslinking analysis of preferential interactions between a transmembrane peptide and matching lipids

Ridder, A.N.J.A., Spelbrink, R.E.J., Demmers, J.A.A., Rijkers, D.T.S., Liskamp, R.M.J. , Brunner, J., Heck, A.J.R., de Kruijff, B. and Killian, J.A. (2004) Photo-crosslinking analysis of preferential interactions between a transmembrane peptide and matching lipids. Biochemistry, 43(15), pp. 4482-4489. (doi:10.1021/bi049899d)

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Abstract

n this study, a novel method is presented by which the molecular environment of a transmembrane peptide can be investigated directly. This was achieved by incorporating a photoactivatable crosslinking probe in the hydrophobic segment of a model transmembrane peptide. When this peptide was incorporated into lipid bilayers and irradiated with UV light, a covalent bond was formed between the crosslinking probe and a lipid. This crosslinking reaction could be visualized by sodium dodecyl sulfate−polyacrylamide gel electrophoresis and the resulting product could be characterized by mass spectrometry. By use of phospholipases, it was demonstrated that the peptide crosslinks to both acyl chains of the lipids. The peptide showed a clear preference to partition into fluid lipids and was excluded from lipids in the gel phase. However, when the peptide was incorporated into bilayers containing two lipid species with different acyl chain lengths, molecular sorting of the lipids around the peptide based on hydrophobic matching was not observed. It is proposed that the size of the transmembrane part plays an important role in the dynamic interactions of membrane proteins with the surrounding lipids and hence in determining whether molecular sorting can occur.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Liskamp, Professor Robert
Authors: Ridder, A.N.J.A., Spelbrink, R.E.J., Demmers, J.A.A., Rijkers, D.T.S., Liskamp, R.M.J., Brunner, J., Heck, A.J.R., de Kruijff, B., and Killian, J.A.
College/School:College of Science and Engineering > School of Chemistry
Journal Name:Biochemistry
Publisher:American Chemical Society
ISSN:0006-2960
ISSN (Online):1520-4995

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