Interaction between biotin lipids and streptavidin in monolayers: formation of oriented two-dimensional protein domains induced by surface recognition

Blankenburg, R.; Meller, P.; Ringsdorf, Helmut; Salesse, Christian

doi:10.1021/bi00446a037

Cited by 253 publications

(190 citation statements)

References 25 publications

(16 reference statements)

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“…In many cases, closely packed arrays or twodimensional crystals have been found to facilitate attaining high resolution (39 -41). One strategy that has been explored to obtain such arrays is the use of streptavidin films grown on biotinylated lipid films (42) as a matrix for growing oriented arrays of biotinylated proteins (43,44). However, the selforganization of the underlying two-dimensional crystalline streptavidin matrix imposes severe steric limitations on the two-dimensional arrangement of proteins bound to it and, moreover, these proteins need to be biotinylated in a sitespecific manner.…”

Section: Binding Of His-tagged Proteins To Metal-chelating Lipidmentioning

confidence: 99%

Specific Orientation and Two-dimensional Crystallization of the Proteasome at Metal-chelating Lipid Interfaces

Theß¹,

Hutschenreiter²,

Hofmann³

et al. 2002

Journal of Biological Chemistry

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The potential of a protein-engineered His tag to immobilize macromolecules in a predictable orientation at metal-chelating lipid interfaces was investigated using recombinant 20 S proteasomes His-tagged in various positions. Electron micrographs demonstrated that the orientation of proteasomes bound to chelating lipid films could be controlled via the location of their His tags: proteasomes His-tagged at their sides displayed exclusively side-on views, while proteasomes His-tagged at their ends displayed exclusively end-on views. The activity of proteasomes immobilized at chelating lipid interfaces was well preserved. In solution, His-tagged proteasomes hydrolyzed casein at rates comparable with wild-type proteasomes, unless the His tags were located in the vicinity of the N termini of ␣-subunits. The N termini of ␣-subunits might partly occlude the entrance channel in ␣-rings through which substrates enter the proteasome for subsequent degradation. A combination of electron micrographs and atomic force microscope topographs revealed a propensity of vertically oriented proteasomes to crystallize in two dimensions on fluid lipid films. The oriented immobilization of His-tagged proteins at biocompatible lipid interfaces will assist structural studies as well as the investigation of biomolecular interaction via a wide variety of surface-sensitive techniques including single-molecule analysis.

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Section: Binding Of His-tagged Proteins To Metal-chelating Lipidmentioning

confidence: 99%

Specific Orientation and Two-dimensional Crystallization of the Proteasome at Metal-chelating Lipid Interfaces

Theß¹,

Hutschenreiter²,

Hofmann³

et al. 2002

Journal of Biological Chemistry

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“…18 Streptavidin is a protein that is comprised of four identical subunits, each binding one biotin molecule. The binding affinity between streptavidin and biotin is so high (k R ) 10 15 M -1 ) that the formation of this complex can be regarded as nearly irreversible, on a scare nearly comparable to a covalent bond.…”

Section: Introductionmentioning

confidence: 99%

Oriented Assembly of Purple Membrane on Solid Support, Mediated by Molecular Recognition

Chen

Sui

et al. 2003

J. Phys. Chem. B

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Oriented films of purple membrane patches were assembled on solid supports by the biotin/streptavidin molecular recognition technique. The degree of biotin labeling was 0.58/bacteriorhodopsin. According to the kinetics of absorption change after flash illumination, the photochemical activity of the purple membrane was hardly affected by the biotinylation. The three steps of the oriented assembly were visualized by electron microscopy. According to surface plasmon resonance, about 0.46 ng/mm 2 of streptavidin and 1.16 ng/mm 2 of biotinylated purple membrane were in the assembled films. The amount of the nonspecifically adsorbed purple membrane, without label, was 0.44 ng/mm 2 . To test the properties of this oriented purple membrane film, a similar assembly procedure was carried out with the bilayer lipid membrane system. The photoelectric voltage peak of the oriented purple membrane was 157.2 mV, whereas that of nonspecific adsorbed purple membrane was 7.8 mV. The degree of orientation was calculated to be 81% theoretically. Further washing experiments suggested that the nonspecific adsorption may be attributed to both the electrostatic attraction and the hydrophobic interaction.

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“…The extremely tight biotin-binding ability of streptavidin not only offers useful bioanalytical applications (4,5) but also generates considerable protein chemical interest, particularly as an attractive model for studying macromolecule-ligand interactions (6)(7)(8)(9)(10)(11)(12)(13). One such effort was made by determining the three-dimensional structure of streptavidin by x-ray crystallography (14,15).…”

mentioning

confidence: 99%

Intersubunit contacts made by tryptophan 120 with biotin are essential for both strong biotin binding and biotin-induced tighter subunit association of streptavidin.

Sano

Cantor

1995

Proc. Natl. Acad. Sci. U.S.A.

132

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BiochemistryIntersubunit contacts made by tryptophan 120 with biotin are essential for both strong biotin binding and biotin-induced tighter subunit association of streptavidin (protein ABSTRACTIn natural streptavidin, tryptophan 120 of each subunit makes contacts with the biotin bound by an adjacent subunit through the dimer-dimer interface. To understand quantitatively the role of tryptophan 120 and its intersubunit communication in the properties of streptavidin, a streptavidin mutant in which tryptophan 120 is converted to phenylalanine was produced and characterized. The streptavidin mutant forms a tetrameric molecule and binds one biotin per subunit, as does natural streptavidin, indicating that the mutation of tryptophan 120 to phenylalanine has no significant effect on the basic properties of streptavidin. However, its biotin-binding affinity was reduced substantially, to approximately 108 M-l, indicating that the contact made by tryptophan 120 to biotin has a considerable contribution to the extremely tight biotin binding by streptavidin. The mutant retained bound biotin over a wide pH range or with the addition of urea up to 6 M at neutral pH. However, bound biotin was efficiently released by the addition of excess free biotin due, presumably, to exchange reactions. Electrophoretic analysis revealed that the intersubunit contact made by tryptophan 120 to biotin through the dimer-dimer interface is the major interaction responsible for the biotininduced, tighter subunit association of streptavidin. In addition, the mutant has weaker subunit association than natural streptavidin even in the absence of biotin, indicating that tryptophan 120 also contributes to the subunit association of tetramers in the absence of biotin.

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Interaction between biotin lipids and streptavidin in monolayers: formation of oriented two-dimensional protein domains induced by surface recognition

Cited by 253 publications

References 25 publications

Specific Orientation and Two-dimensional Crystallization of the Proteasome at Metal-chelating Lipid Interfaces

Specific Orientation and Two-dimensional Crystallization of the Proteasome at Metal-chelating Lipid Interfaces

Oriented Assembly of Purple Membrane on Solid Support, Mediated by Molecular Recognition

Intersubunit contacts made by tryptophan 120 with biotin are essential for both strong biotin binding and biotin-induced tighter subunit association of streptavidin.

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