Molecular organization of histidine-tagged biomolecules at self-assembled lipid interfaces using a novel class of chelator lipids.
In molecular biology, the expression of fusion proteins is a very useful and well-established technique for the identification and one-step purification of gene products. Even a short fused sequence of five or six histidines enables proteins to bind to an immobilized metal ion chelate complex. By synthesis of a class of chelator lipids, we have transferred this approach to the concept of self-assembly. The specific interaction and lateral organization of a fluorescent fusion molecule containing a C-terminal oligohistidine sequence was studied by film balance techniques in combination with epifluorescence microscopy. Due to the phase behavior of the various lipid mixtures used, the chelator lipids can be laterally structured, generating two-dimensional arrays of histidine-tagged biomolecules. Because of the large variety of fusion proteins already available, this concept represents a powerful technique for orientation and organization of proteins at lipid interfaces with applications in biosensing, biofunctionalization of nanostructured interfaces, two-dimensional crystallization, and studies of lipid-anchored proteins.
Dietrich
C
C
Lehrstuhl für Biophysik E22, Technische Universität München, Garching, Germany.
Schmitt
L
L
Tampé
R
R
D000588:Amines / Q000737:chemistry*; D000595:Amino Acid Sequence; D002614:Chelating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D004134:Dimyristoylphosphatidylcholine; D006639:Histidine; D007202:Indicators and Reagents; D008961:Models, Structural; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D016324:Sequence Tagged Sites
D000588:Amines; D002614:Chelating Agents; D007202:Indicators and Reagents; D010455:Peptides; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins; C095818:nitrilotriacetic acid dioctadecylamine; C095819:dioctadecylamine; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine; D006639:Histidine; D009571:Nitrilotriacetic Acid; D004134:Dimyristoylphosphatidylcholine