Extremely high thermal stability of streptavidin and avidin upon biotin binding

González, Martı́n; Argaraña, Carlos E.; Fidelio, Gerardo D.

doi:10.1016/s1050-3862(99)00041-8

Cited by 170 publications

(151 citation statements)

References 20 publications

Supporting

Mentioning

137

Contrasting

Order By: Relevance

“…S2). These results are consistent with the well characterized stabilizing effect of biotin on avidin (35,38) and show the possibility for achieving microstructure actuation by means of biotin binding. By using engineered avidins (37,39), biotininduced actuation can be explored under a wider range of environmental conditions.…”

Section: Resultssupporting

confidence: 88%

Multiphoton fabrication of chemically responsive protein hydrogels for microactuation

Kaehr

Shear

2008

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

182

181

View full text Add to dashboard Cite

We report a method for creating stimuli-responsive biomaterials in which scanning nonlinear excitation is used to photocrosslink proteins at submicrometer 3D coordinates. Proteins with differing hydration properties can be combined to achieve tunable volume changes that are rapid and reversible in response to changes in chemical environment. Protein matrices having arbitrary 3D topographies and definable density gradients over micrometer dimensions provide the ability to effect rapid (<1 sec) and precise mechanical manipulations by means of changes in hydrogel size and shape, and applicability of these materials to cell biology is shown through the fabrication of responsive bacterial cages.Escherichia coli ͉ multiphoton lithography ͉ nanobiotechnology ͉ protein hydrogels ͉ smart materials

show abstract

Section: Resultssupporting

confidence: 88%

Multiphoton fabrication of chemically responsive protein hydrogels for microactuation

Kaehr

Shear

2008

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

182

181

View full text Add to dashboard Cite

show abstract

“…ues for rhizavidin and other avidins (6,45), clearly validating shwanavidin as a thermostable protein ( Table 2).…”

Section: Resultsmentioning

confidence: 71%

Structural Adaptation of a Thermostable Biotin-binding Protein in a Psychrophilic Environment

Meir

Bayer

Livnah

2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: Search for novel avidins with reduced oligomeric state revealed an intriguing dimeric protein.Results: Structures of shwanavidin from Shewanella denitrificans identified features accountable for high affinity and psychrophilic properties. Conclusion: Phe-43 and disulfide bridge are responsible for the high affinity compensating the lack of intermonomeric interaction in tetrameric avidins. Significance: Proteins from extremophile organisms provide excellent platforms toward the development of novel biotechnological utilizations.

show abstract

“…Weaker subunit interaction would result in a higher percentage of the sample in the monomeric state on the SDSpolyacrylamide gel. Because biotin can strengthen subunit interaction, samples were analyzed in the presence or absence of biotin (9,26). The impact of the mutation on weakening of the subunit interaction followed the order: T76R Ͼ V125R Ͼ V125T Ϸ V55R Ͼ V55T (Fig.…”

Section: Effects Of Single Mutations On Monomerization Ofmentioning

confidence: 99%

Engineering Soluble Monomeric Streptavidin with Reversible Biotin Binding Capability

Wong

2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Monomeric streptavidin with reversible biotin binding capability has many potential applications. Because a complete biotin binding site in each streptavidin subunit requires the contribution of tryptophan 120 from a neighboring subunit, monomerization of the natural tetrameric streptavidin can generate streptavidin with reduced biotin binding affinity. Three residues, valine 55, threonine 76, and valine 125, were changed to either arginine or threonine to create electrostatic repulsion and steric hindrance at the interfaces. The double mutation (T76R,V125R) was highly effective to monomerize streptavidin. Because interfacial hydrophobic residues are exposed to solvent once tetrameric streptavidin is converted to the monomeric state, a quadruple mutein (T76R,V125R,V55T,L109T) was developed. The first two mutations are for monomerization, whereas the last two mutations aim to improve hydrophilicity at the interface to minimize aggregation. Monomerization was confirmed by four different approaches including gel filtration, dynamic light scattering, sensitivity to proteinase K, and chemical cross-linking. The quadruple mutein remained in the monomeric state at a concentration greater than 2 mg/ml. Its kinetic parameters for interaction with biotin suggest excellent reversible biotin binding capability, which enables the mutein to be easily purified on the biotin-agarose matrix. Another mutein (D61A,W120K) was developed based on two mutations that have been shown to be effective in monomerizing avidin. This streptavidin mutein was oligomeric in nature. This illustrates the importance in selecting the appropriate residues and approaches for effective monomerization of streptavidin.(Strept)avidin with reversible biotin binding capability can extend the applications of the biotin-(strept)avidin technology. These molecules can be applied for affinity purification of biotinylated biomolecules, screening of ultratight binders binding to biotinylated biomolecules displayed on the phage display system, and development of reusable biosensor chips, protein/ antibody microarrays, and enzyme bioreactors (1, 2). (Strept)avidin is a homotetrameric molecule with a biotin binding site in each subunit (3). The three-dimensional structure of (strept)avidin (4, 5) suggests that a complete biotin binding pocket in each subunit requires the contribution of a tryptophan residue from an adjacent subunit. Site-directed mutagenesis studies also demonstrate the importance of this residue for tight biotin binding and subunit communications (6 -8). Therefore, development of monomeric (strept)avidin can be an attractive approach to engineer (strept)avidin with reversible biotin binding capability.The engineering of (strept)avidin to its monomeric form is technically challenging. In the case of avidin, the first generation of engineered monomeric avidin can exist in the monomeric state only in the absence of biotin (9). This problem has been solved by the recent development of the second generation of monomeric avidin (10), which carries two mut...

show abstract

Extremely high thermal stability of streptavidin and avidin upon biotin binding

Cited by 170 publications

References 20 publications

Multiphoton fabrication of chemically responsive protein hydrogels for microactuation

Multiphoton fabrication of chemically responsive protein hydrogels for microactuation

Structural Adaptation of a Thermostable Biotin-binding Protein in a Psychrophilic Environment

Engineering Soluble Monomeric Streptavidin with Reversible Biotin Binding Capability

Contact Info

Product

Resources

About