Engineering Two-Dimensional Protein Order at Surfaces

Koppenol, Sandy; Stayton, Patrick S.

doi:10.1021/js970067t

Cited by 6 publications

(9 citation statements)

References 88 publications

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“…27 The presence of two pairs of biotin-binding sites on opposing sides of each streptavidin molecule, along with its high affinity for biotin, results in extensive use as a molecular linker in various applications. [28][29][30] This work aims toward a fundamental understanding of self-assembly processes in biological systems. It has been known that hundreds of species of bacteria and archea found in various environments contain a crystalline layer of surface proteins at the cell surface called s-layers.…”

Section: Introductionmentioning

confidence: 99%

Mechanics of Streptavidin-Coated Giant Lipid Bilayer Vesicles: A Micropipet Study

et al. 2003

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To understand the effects of a crystalline protein layer on bilayer properties, we studied the mechanical properties of avidin-and streptavidin-coated giant lipid bilayer vesicles. The giant vesicles (20-60 µm) are made from a mixture of SOPC and biotinylated phospholipids via electroformation. Using micropipet manipulation, we showed that the presence of a monomolecular layer of noncrystalline avidin on the vesicle surface increases the membrane bending rigidity but does not significantly alter the elastic area expansion modulus of the vesicle. When the vesicles were coated with streptavidin, the protein crystallizes on the bilayer surface, resulting in a rigid polycrystalline membrane. These vesicles display unique roughened spherical or prolate ellipsoidal shapes, depending on the differences in crystal morphologies. Upon aspiration with micropipets, the vesicles first showed rapid permanent deformation at low strain, followed by a slower viscoelastic response above a certain threshold. Despite their extremely rigid appearance, the existence of a polycrystalline shell does not increase the toughness of streptavidin-coated vesicles above that of uncoated vesicles. The origin of these properties can be traced to the unique ligand-receptor interactions between streptavidin and biotinylated phospholipids in the bilayer membrane. The findings offer greater understandings of complex phenomena involving crystalline protein layers on the surface of cell membranes, in addition to providing information for the development of various applications involving the immobilization of functionalized molecules on lipid bilayer substrates.

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Section: Introductionmentioning

confidence: 99%

Mechanics of Streptavidin-Coated Giant Lipid Bilayer Vesicles: A Micropipet Study

et al. 2003

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“…[4][5][6][7] The ability for streptavidin to crystallize into well-ordered arrays creates potential for numerous applications in the field of biosensors and biomaterials. [8][9][10] The fact that streptavidin molecules contain four biotin binding sites, two pairs on opposite side of the molecules, 6 makes them ideal as strong cross-linkers. Due to the unusually high binding affinity for biotin (10 -15 M), 5 the streptavidin-biotin system has been widely exploited for various physical and biological applications.…”

Section: Introductionmentioning

confidence: 99%

Two-Dimensional Streptavidin Crystals on Giant Lipid Bilayer Vesicles

et al. 2002

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Streptavidin was crystallized on giant bilayer vesicles (20-60 µm) in sucrose solution at various pH values. The streptavidin-coated vesicles exhibited unique roughened spherical and prolate ellipsoidal shapes, illustrating resistance to curvature of the two-dimensional crystals. Studies indicated that the spheroids and prolate ellipsoids correspond to different crystal morphologies. Through confocal microscopy, the various crystal morphologies on vesicle surfaces were observed under different solution conditions. Unlike two-dimensional (2D) streptavidin crystals grown in ionic buffer that assume the P1, P2, and C222 lattices at pH 4, 5.5, and 7, respectively (Wang et al. Langmuir 1999, 15, 1541), crystals grown in sucrose with no added salt show only the lowest density C222 lattice due to strong electrostatic interactions.

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“…This information is needed to grow larger, more ordered crystals for protein structure analysis. 20,21 Additionally, this work has implications in the development of interfacial technologies such as biosensors and biomaterials, [22][23][24] which require presentation of a protein binding site or functional group in a highly ordered and oriented configuration.…”

Section: Introductionmentioning

confidence: 99%

“…Another application of the streptavidin−biotin system encompasses the study of self-assembling protein arrays. − Streptavidin molecules bound to a monolayer of biotinylated lipid at an air−water interface can form two-dimensional (2D) protein crystals. ,, We use this system to examine interactions among proteins to obtain a better understanding of the effects they have on 2D self-assembly phenomena. This information is needed to grow larger, more ordered crystals for protein structure analysis. , Additionally, this work has implications in the development of interfacial technologies such as biosensors and biomaterials, − which require presentation of a protein binding site or functional group in a highly ordered and oriented configuration.…”

Section: Introductionmentioning

confidence: 99%

Molecular Arrangement in Two-Dimensional Streptavidin Crystals

1999

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We studied the molecular arrangement of two-dimensional streptavidin crystals at the air-water interface over a range of pH values. We quantified the varying amounts of coexisting P1, P2, and C222 crystals in the different morphologies observed at pH 4.5-6.5. Chiral, needlelike crystals at pH 4.5 consist of P1 crystals with frequent line defects. Larger chiral domains near pH 5 are essentially all P1 coexisting with a small amount of P2, whereas at slightly higher pH values (near pH 5.5), H-shaped domains contain 4 times as much P1 coexisting with a P2/C222 mixture. Morphologies intermediate to these shapes exhibit intermediate compositions. Between pH ∼6-7, crystals all display a characteristic dendritic-X morphology, but arrangement at the molecular level is quite different compared with lower pH values. Crystals are mostly P2 in symmetry near pH 6, but at pH 7 and above, crystals have C222 symmetry. Coexistence of P2 and C222 crystals occurs at intermediate pH values. We determined the orientation and arrangement of streptavidin molecules in P1, P2, and C222 crystals relative to the directions exhibiting faster growth. The direction of faster growth in P1 crystals includes both interactions between biotin-free subunits and interactions between biotin-bound subunits. In the P2 arrangement, growth in the direction of intermolecular biotin-free subunits is preferred, whereas growth is faster along the biotin-bound direction of C222 crystals. We developed a model of the molecular arrangement for the observed solid-phase coexistence in these crystals.

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Engineering Two-Dimensional Protein Order at Surfaces

Cited by 6 publications

References 88 publications

Mechanics of Streptavidin-Coated Giant Lipid Bilayer Vesicles: A Micropipet Study

Mechanics of Streptavidin-Coated Giant Lipid Bilayer Vesicles: A Micropipet Study

Two-Dimensional Streptavidin Crystals on Giant Lipid Bilayer Vesicles

Molecular Arrangement in Two-Dimensional Streptavidin Crystals

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