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首页> 外文期刊>Biochemistry >Characterization of Self-Association and Heteroassociation of Bacterial Cell Division Proteins FtsZ and ZipA in Solution by Composition Gradient−Static Light Scattering
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Characterization of Self-Association and Heteroassociation of Bacterial Cell Division Proteins FtsZ and ZipA in Solution by Composition Gradient−Static Light Scattering

机译：组成梯度梯度静态光散射表征溶液中细菌细胞分裂蛋白FtsZ和ZipA的自缔合和杂缔合

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We have characterized the self-association of FtsZ in its GDP-bound state (GDP-FtsZ) and thenheteroassociation of FtsZ and a soluble recombinant ZipA (sZipA) lacking the N-terminal transmembranendomain by means of composition gradient-static light scattering (CG-SLS) and by measurement ofnsedimentation equilibrium. CG-SLS experiments at high ionic strengths and in the presence of 5 mM Mg2þnshow that, while FtsZ self-associates in a noncooperative fashion, sZipA acts as a monomer. CG-SLS datanobtained from mixtures of FtsZ (A) and sZipA (B) in the presence ofMg2þ are quantitatively described by annequilibrium model that takes into account significant scattering contributions from B, A1,A2,A3,A4,A5,A6,nA1B, A2B, A3B, and A4B. However, in the absence of Mg2þ (with EDTA), the data are best explained by annequilibrium model in which only B, A1,A2,A3,A1B, and A2B contribute significantly to scattering. The best-nfit molecular weights of monomeric A and B are in good agreement with values calculated from amino acidncomposition and with values obtained from sedimentation equilibrium. The latter technique also confirmednthe interaction between sZipA and GDP-FtsZ. Moreover, the association model that best describes thenCG-SLS data is in qualitative agreement with the sedimentation data. From these results, it follows that thenbinding of sZipA to GDP-FtsZ is of moderate affinity and does not significantly affect the interactionsnbetween FtsZ monomers. Under the working conditions used, only one sZipA binds to FtsZ oligomers with anlength of six at most. The observed behavior would be compatible with FtsZ fibrils being anchored in vivo tonthe bacterial inner plasma membrane by substoichiometric binding of membrane-bound ZipA.
机译：我们已经表征了FtsZ在其GDP结合状态（GDP-FtsZ）的自缔合，然后通过组成梯度静态光散射（CG-）的方式将FtsZ与缺少N端跨膜结构域的可溶性重组ZipA（sZipA）进行异缔合。 SLS），并通过测量沉降平衡。 CG-SLS在高离子强度和5 mM Mg2 + n存在下的实验表明，尽管FtsZ以非合作方式自缔合，但sZipA却是单体。在Mg2 +存在下从FtsZ（A）和sZipA（B）的混合物中获得的CG-SLS数据通过平衡模型定量描述，该模型考虑了B，A1，A2，A3，A4，A5，A6，nA1B， A2B，A3B和A4B。但是，在不存在Mg2 +（使用EDTA）的情况下，最好通过退火模型解释数据，其中只有B，A1，A2，A3，A1B和A2B对散射有显着贡献。单体A和B的最佳拟合分子量与从氨基酸组成计算得出的值以及从沉降平衡获得的值非常吻合。后一种技术也证实了sZipA与GDP-FtsZ之间的相互作用。此外，最能描述CG-SLS数据的关联模型与沉积数据在质量上是一致的。从这些结果可以看出，sZipA与GDP-FtsZ的结合具有中等亲和力，并且不会显着影响FtsZ单体之间的相互作用。在所使用的工作条件下，只有一个sZipA结合至FtsZ寡聚体的长度最多为六个。所观察到的行为将与通过膜结合的ZipA的亚化学计量结合将FtsZ原纤维在体内锚定在细菌内质膜上相容。

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  • 来源
    《Biochemistry》 |2010年第51期|p.10780-10787|共8页
  • 作者

    Ariadna Martos Carlos Alfonso Pilar Lpez-Navajas Rubn Ahijado-Guzmn Jess Mingorance Allen P. Minton and Germn Rivas;

  • 作者单位

    ‡Chemical and Physical Biology Program, Centro de Investigaciones Biolu0001 ogicas, and §Department of Molecular Biotechnology,Centro Nacional de Biotecnologı ´a, Consejo Superior de Investigaciones Cientı ´ficas, Madrid, Spain, and ) Section on PhysicalBiochemistry, Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases,National Institutes of Health, Bethesda,Maryland 20892, United States.^Current address: Servicio de Microbiologı ´a,Hospital Universitario La Paz, IdiPAZ, Madrid, Spain.;

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