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

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Freitag, S; Le Trong, I; Klumb, LA; Chu, V; Chilkoti, A; Stayton, PS; Stenkamp, RE

Published Date

  • December 1999

Published In

Volume / Issue

  • 16 / 1-4

Start / End Page

  • 13 - 19

PubMed ID

  • 10796980

International Standard Serial Number (ISSN)

  • 1389-0344

Digital Object Identifier (DOI)

  • 10.1016/s1050-3862(99)00048-0


  • eng