1n4j: Difference between revisions
New page: left|200px<br /><applet load="1n4j" size="450" color="white" frame="true" align="right" spinBox="true" caption="1n4j, resolution 2.18Å" /> '''STREPTAVIDIN MUTANT ... |
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[[Image:1n4j.gif|left|200px]]<br /><applet load="1n4j" size=" | [[Image:1n4j.gif|left|200px]]<br /><applet load="1n4j" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1n4j, resolution 2.18Å" /> | caption="1n4j, resolution 2.18Å" /> | ||
'''STREPTAVIDIN MUTANT N23A AT 2.18A'''<br /> | '''STREPTAVIDIN MUTANT N23A AT 2.18A'''<br /> | ||
==Overview== | ==Overview== | ||
An elaborate hydrogen-bonding network contributes to the tight binding of | 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. | ||
==About this Structure== | ==About this Structure== | ||
1N4J is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_avidinii Streptomyces avidinii]. Full crystallographic information is available from [http:// | 1N4J is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_avidinii Streptomyces avidinii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1N4J OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Chu, V.]] | [[Category: Chu, V.]] | ||
[[Category: Freitag, S.]] | [[Category: Freitag, S.]] | ||
[[Category: Klumb, L | [[Category: Klumb, L A.]] | ||
[[Category: Stayton, P | [[Category: Stayton, P S.]] | ||
[[Category: Stenkamp, R | [[Category: Stenkamp, R E.]] | ||
[[Category: Trong, I | [[Category: Trong, I Le.]] | ||
[[Category: homotetramer]] | [[Category: homotetramer]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:02:10 2008'' | ||
Revision as of 15:02, 21 February 2008
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STREPTAVIDIN MUTANT N23A AT 2.18A
OverviewOverview
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.
About this StructureAbout this Structure
1N4J is a Single protein structure of sequence from Streptomyces avidinii. Full crystallographic information is available from OCA.
ReferenceReference
Structural studies of hydrogen bonds in the high-affinity streptavidin-biotin complex: mutations of amino acids interacting with the ureido oxygen of biotin., Le Trong I, Freitag S, Klumb LA, Chu V, Stayton PS, Stenkamp RE, Acta Crystallogr D Biol Crystallogr. 2003 Sep;59(Pt 9):1567-73. Epub 2003, Aug 19. PMID:12925786
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