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X-ray crystallographic studies of streptavidin mutants binding to biotin.

Publication ,  Journal Article
Freitag, S; Le Trong, I; Klumb, LA; Chu, V; Chilkoti, A; Stayton, PS; Stenkamp, RE
Published in: Biomolecular engineering
December 1999

On the basis of high resolution crystallographic studies of streptavidin and its biotin complex, three principal binding motifs have been identified that contribute to the tight binding. A flexible binding loop can undergo a conformational change from an open to a closed form when biotin is bound. Additional studies described here of unbound wild-type streptavidin have provided structural views of the open conformation. Several tryptophan residues packing around the bound biotin constitute the second binding motif, one dominated by hydrophobic interactions. Mutation of these residues to alanine or phenylalanine have variable effects on the thermodynamics and kinetics of binding, but they generate only small changes in the molecular structure. Hydrogen bonding interactions also contribute significantly to the binding energetics of biotin, and the D128A mutation which breaks a hydrogen bond between the protein and a ureido NH group results in a significant structural alteration that could mimic an intermediate on the dissociation pathway. In this review, we summarize the structural aspects of biotin recognition that have been gained from crystallographic analyses of wild-type and site-directed streptavidin mutants.

Duke Scholars

Published In

Biomolecular engineering

DOI

ISSN

1389-0344

Publication Date

December 1999

Volume

16

Issue

1-4

Start / End Page

13 / 19

Related Subject Headings

  • Streptavidin
  • Protein Engineering
  • Protein Conformation
  • Point Mutation
  • Mutagenesis, Site-Directed
  • Models, Molecular
  • Hydrogen Bonding
  • Crystallography, X-Ray
  • Biotin
  • Biotechnology
 

Citation

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MLA
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Freitag, S., Le Trong, I., Klumb, L. A., Chu, V., Chilkoti, A., Stayton, P. S., & Stenkamp, R. E. (1999). X-ray crystallographic studies of streptavidin mutants binding to biotin. Biomolecular Engineering, 16(1–4), 13–19. https://doi.org/10.1016/s1050-3862(99)00048-0
Freitag, S., I. Le Trong, L. A. Klumb, V. Chu, A. Chilkoti, P. S. Stayton, and R. E. Stenkamp. “X-ray crystallographic studies of streptavidin mutants binding to biotin.Biomolecular Engineering 16, no. 1–4 (December 1999): 13–19. https://doi.org/10.1016/s1050-3862(99)00048-0.
Freitag S, Le Trong I, Klumb LA, Chu V, Chilkoti A, Stayton PS, et al. X-ray crystallographic studies of streptavidin mutants binding to biotin. Biomolecular engineering. 1999 Dec;16(1–4):13–9.
Freitag, S., et al. “X-ray crystallographic studies of streptavidin mutants binding to biotin.Biomolecular Engineering, vol. 16, no. 1–4, Dec. 1999, pp. 13–19. Epmc, doi:10.1016/s1050-3862(99)00048-0.
Freitag S, Le Trong I, Klumb LA, Chu V, Chilkoti A, Stayton PS, Stenkamp RE. X-ray crystallographic studies of streptavidin mutants binding to biotin. Biomolecular engineering. 1999 Dec;16(1–4):13–19.
Journal cover image

Published In

Biomolecular engineering

DOI

ISSN

1389-0344

Publication Date

December 1999

Volume

16

Issue

1-4

Start / End Page

13 / 19

Related Subject Headings

  • Streptavidin
  • Protein Engineering
  • Protein Conformation
  • Point Mutation
  • Mutagenesis, Site-Directed
  • Models, Molecular
  • Hydrogen Bonding
  • Crystallography, X-Ray
  • Biotin
  • Biotechnology