1N4J
STREPTAVIDIN MUTANT N23A AT 2.18A
Summary for 1N4J
| Entry DOI | 10.2210/pdb1n4j/pdb |
| Related | 1N43 1SWE |
| Descriptor | Streptavidin (2 entities in total) |
| Functional Keywords | homotetramer, biotin-binding protein |
| Biological source | Streptomyces avidinii |
| Cellular location | Secreted: P22629 |
| Total number of polymer chains | 1 |
| Total formula weight | 13238.31 |
| Authors | Le Trong, I.,Freitag, S.,Klumb, L.A.,Chu, V.,Stayton, P.S.,Stenkamp, R.E. (deposition date: 2002-10-31, release date: 2003-09-02, Last modification date: 2024-02-14) |
| Primary citation | Le Trong, I.,Freitag, S.,Klumb, L.A.,Chu, V.,Stayton, P.S.,Stenkamp, R.E. Structural studies of hydrogen bonds in the high-affinity streptavidin-biotin complex: mutations of amino acids interacting with the ureido oxygen of biotin. Acta Crystallogr.,Sect.D, 59:1567-1573, 2003 Cited by PubMed Abstract: An elaborate hydrogen-bonding network contributes to the tight binding of biotin to streptavidin. The specific energetic contributions of hydrogen bonds to the biotin ureido oxygen have previously been investigated by mapping the equilibrium and activation thermodynamic signatures of N23A, N23E, S27A, Y43A and Y43F site-directed mutants [Klumb et al. (1998), Biochemistry, 37, 7657-7663]. The crystal structures of these variants in the unbound and biotin-bound states provide structural insight into the energetic alterations and are described here. High (1.5-2.2 A) to atomic resolution (1.14 A) structures were obtained and structural models were refined to R values ranging from 0.12 to 0.20. The overall folding of streptavidin as described previously has not changed in any of the mutant structures. Major deviations such as side-chain shifts of residues in the binding site are observed only for the N23A and Y43A mutations. In none of the mutants is a systematic shift of biotin observed when one of the hydrogen-bonding partners to the ureido oxygen of biotin is removed. Recent thermodynamic studies report increases of DeltaDeltaG(o) of 5.0-14.6 kJ mol(-1) for these mutants with respect to the wild-type protein. The decreasing stabilities of the complexes of the mutants are discussed in terms of their structures. PubMed: 12925786DOI: 10.1107/S0907444903014562 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.18 Å) |
Structure validation
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