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Large conformational changes in the Escherichia coli tryptophan synthase beta(2) subunit upon pyridoxal 5'-phosphate binding.
FEBS J. 2010 May; 277(9):2157-70.FJ

Abstract

To understand the basis for the lower activity of the tryptophan synthase beta(2) subunit in comparison to the alpha(2)beta(2) complex, we determined the crystal structures of apo-beta(2) and holo-beta(2) from Escherichia coli at 3.0 and 2.9 A resolutions, respectively. To our knowledge, this is the first report of both beta(2) subunit structures with and without pyridoxal-5'-phosphate. The apo-type molecule retained a dimeric form in solution, as in the case of the holo-beta(2) subunit. The subunit structures of both the apo-beta(2) and the holo-beta(2) forms consisted of two domains, namely the N domain and the C domain. Although there were significant structural differences between the apo- and holo-structures, they could be easily superimposed with a 22 degrees rigid body rotation of the C domain. The pyridoxal-5'-phosphate-bound holo-form had multiple interactions between the two domains and a long loop (residues 260-310), which were missing in the apo-form. Comparison of the structures of holo-Ecbeta(2) and Stbeta(2) in the alpha(2)beta(2) complex from Salmonella typhimurium (Stalpha(2)beta(2)) identified the cause of the lower enzymatic activity of holo-Ecbeta(2) in comparison with Stalpha(2)beta(2). The substrate (indole) gate residues, Tyr279 and Phe280, block entry of the substrate into the beta(2) subunit, although the indole can directly access the active site as a result of a wider cleft between the N and C domains in the holo-Ecbeta(2) subunit. In addition, the structure around betaAsp305 of the holo-Ecbeta(2) subunit was similar to the open state of Stalpha(2)beta(2) with low activity, resulting in lower activity of holo-Ecbeta(2).

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Authors+Show Affiliations

Nishio K
Institute for Protein Research, Osaka University, Japan.
Ogasahara K
No affiliation info available
Morimoto Y
No affiliation info available
Tsukihara T
No affiliation info available
Lee SJ
No affiliation info available
Yutani K
No affiliation info available

MeSH

Allosteric SiteAmino Acid SequenceApoenzymesCrystallography, X-RayEnzyme StabilityEscherichia coliHoloenzymesHydrogen-Ion ConcentrationModels, MolecularMolecular Sequence DataProtein MultimerizationProtein Structure, QuaternaryProtein SubunitsPyridoxal PhosphateSalmonella typhimuriumSequence AlignmentTryptophan Synthase

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20370823

Citation

Nishio, Kazuya, et al. "Large Conformational Changes in the Escherichia Coli Tryptophan Synthase Beta(2) Subunit Upon Pyridoxal 5'-phosphate Binding." The FEBS Journal, vol. 277, no. 9, 2010, pp. 2157-70.
Nishio K, Ogasahara K, Morimoto Y, et al. Large conformational changes in the Escherichia coli tryptophan synthase beta(2) subunit upon pyridoxal 5'-phosphate binding. FEBS J. 2010;277(9):2157-70.
Nishio, K., Ogasahara, K., Morimoto, Y., Tsukihara, T., Lee, S. J., & Yutani, K. (2010). Large conformational changes in the Escherichia coli tryptophan synthase beta(2) subunit upon pyridoxal 5'-phosphate binding. The FEBS Journal, 277(9), 2157-70. https://doi.org/10.1111/j.1742-4658.2010.07631.x
Nishio K, et al. Large Conformational Changes in the Escherichia Coli Tryptophan Synthase Beta(2) Subunit Upon Pyridoxal 5'-phosphate Binding. FEBS J. 2010;277(9):2157-70. PubMed PMID: 20370823.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Large conformational changes in the Escherichia coli tryptophan synthase beta(2) subunit upon pyridoxal 5'-phosphate binding. AU - Nishio,Kazuya, AU - Ogasahara,Kyoko, AU - Morimoto,Yukio, AU - Tsukihara,Tomitake, AU - Lee,Soo Jae, AU - Yutani,Katsuhide, Y1 - 2010/03/27/ PY - 2010/4/8/entrez PY - 2010/4/8/pubmed PY - 2010/5/6/medline SP - 2157 EP - 70 JF - The FEBS journal JO - FEBS J VL - 277 IS - 9 N2 - To understand the basis for the lower activity of the tryptophan synthase beta(2) subunit in comparison to the alpha(2)beta(2) complex, we determined the crystal structures of apo-beta(2) and holo-beta(2) from Escherichia coli at 3.0 and 2.9 A resolutions, respectively. To our knowledge, this is the first report of both beta(2) subunit structures with and without pyridoxal-5'-phosphate. The apo-type molecule retained a dimeric form in solution, as in the case of the holo-beta(2) subunit. The subunit structures of both the apo-beta(2) and the holo-beta(2) forms consisted of two domains, namely the N domain and the C domain. Although there were significant structural differences between the apo- and holo-structures, they could be easily superimposed with a 22 degrees rigid body rotation of the C domain. The pyridoxal-5'-phosphate-bound holo-form had multiple interactions between the two domains and a long loop (residues 260-310), which were missing in the apo-form. Comparison of the structures of holo-Ecbeta(2) and Stbeta(2) in the alpha(2)beta(2) complex from Salmonella typhimurium (Stalpha(2)beta(2)) identified the cause of the lower enzymatic activity of holo-Ecbeta(2) in comparison with Stalpha(2)beta(2). The substrate (indole) gate residues, Tyr279 and Phe280, block entry of the substrate into the beta(2) subunit, although the indole can directly access the active site as a result of a wider cleft between the N and C domains in the holo-Ecbeta(2) subunit. In addition, the structure around betaAsp305 of the holo-Ecbeta(2) subunit was similar to the open state of Stalpha(2)beta(2) with low activity, resulting in lower activity of holo-Ecbeta(2). SN - 1742-4658 UR - https://www.unboundmedicine.com/medline/citation/20370823/Large_conformational_changes_in_the_Escherichia_coli_tryptophan_synthase_beta_2__subunit_upon_pyridoxal_5'_phosphate_binding_ L2 - https://doi.org/10.1111/j.1742-4658.2010.07631.x DB - PRIME DP - Unbound Medicine ER -
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