Farnesylation of human guanylate‐binding protein 1 as safety mechanism preventing structural rearrangements and uninduced dimerization
Farnesylation of human guanylate‐binding protein 1 as safety mechanism preventing structural rearrangements and uninduced dimerization
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人鸟苷酸结合蛋白 1 的法呢基化作为防止结构重排和非诱导二聚化的安全机制
DOI：
10.1111/febs.15015
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Andreas M. Stadler
中科院分区：
文献类型：
--
作者：
Charlotte Lorenz;Semra Ince;Tao Zhang;Anneliese Cousin;Renu Batra-Safferling;Luitgard Nagel-Steger;Christian Herrmann;Andreas M. Stadler
人鸟苷酸结合蛋白1（Human guanylate‐binding protein 1，hGBP 1）是一种动力蛋白（dynamin‐like protein）家族成员，通过添加核苷酸激活，导致蛋白质寡聚化并刺激GTbinding活性。在这项研究中，使用动态光散射（DLS），分析超离心（AUC）和分析尺寸排阻色谱（SEC）研究了farn-hGBP 1和未修饰hGBP 1的流体动力学差异。此外，我们进行了小角X射线散射（SAXS）实验，结合SEC装置（SEC-SAXS），以研究溶液中未修饰的hGBP 1和farn-hGBP 1的结构特性。SEC-SAXS测量显示，法尼基化使hGBP 1在无核苷酸溶液中保持其非活性单体和晶体样构象，而未修饰的hGBP 1在无核苷酸溶液中的非活性基态以及通过添加不可水解的GTP类似物GppNHp捕获的活化状态下形成单体-二聚体平衡。与farn-hGBP相比，未修饰的hGBP 1在结构上受到干扰。特别地，GppNHp结合导致单体和二聚体的大的结构重排和更高的构象灵活性。在未修饰的蛋白质中观察到的结构变化是在核苷酸存在下发生的farn‐ hGBP 1的进一步寡聚体组装的先决条件。数据库所有SEC‐SAXS数据、数据的相应拟合和结构模型都保存在小角散射生物数据库[SASBDB]中。（Nucleic Acids Res，43，2015，D357）]，项目ID为：SASDEE 8、SASDEF 8、SASDEG 8、SASDEH 8、SASDEI 8、SASDEK 8、SASDEL 8和SASDEM 8。
Human guanylate‐binding protein 1 (hGBP1) belongs to the family of dynamin‐like proteins and is activated by addition of nucleotides, leading to protein oligomerization and stimulated GTPase activity.In vivo,hGBP1 is post‐translationally modified by attachment of a farnesyl group yielding farn‐hGBP1. In this study, hydrodynamic differences in farn‐hGBP1 and unmodified hGBP1 were investigated using dynamic light scattering (DLS), analytical ultracentrifugation (AUC) and analytical size‐exclusion chromatography (SEC). In addition, we performed small‐angle X‐ray scattering (SAXS) experiments coupled with a SEC setup (SEC‐SAXS) to investigate structural properties of nonmodified hGBP1 and farn‐hGBP1 in solution. SEC‐SAXS measurements revealed that farnesylation keeps hGBP1 in its inactive monomeric and crystal‐like conformation in nucleotide‐free solution, whereas unmodified hGBP1 forms a monomer–dimer equilibrium both in the inactive ground state in nucleotide‐free solution as well as in the activated state that is trapped by addition of the nonhydrolysable GTP analogue GppNHp. Nonmodified hGBP1 is structurally perturbed as compared to farn‐hGBP. In particular, GppNHp binding leads to large structural rearrangements and higher conformational flexibility of the monomer and the dimer. Structural changes observed in the nonmodified protein are prerequisites for further oligomer assemblies of farn‐hGBP1 that occur in the presence of nucleotides.DatabaseAll SEC‐SAXS data, corresponding fits to the data and structural models are deposited in the Small Angle Scattering Biological Data Bank [SASBDB (Nucleic Acids Res, 43, 2015, D357)] with project IDs: SASDEE8, SASDEF8, SASDEG8, SASDEH8, SASDEJ8, SASDEK8, SASDEL8 and SASDEM8.
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DOI：
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发表时间：
2014
期刊：
影响因子：
--
作者：
M. Rocco;E. Brookes
通讯作者：
E. Brookes
DOI：
10.1107/s1399004714019609
发表时间：
2015-01-01
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
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作者：
De Maria Antolinos A;Pernot P;Brennich ME;Kieffer J;Bowler MW;Delageniere S;Ohlsson S;Malbet Monaco S;Ashton A;Franke D;Svergun D;McSweeney S;Gordon E;Round A
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Round A
影响因子：
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Petoukhov MV;Franke D;Shkumatov AV;Tria G;Kikhney AG;Gajda M;Gorba C;Mertens HD;Konarev PV;Svergun DI
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Svergun DI
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3.9
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Schmid SL
影响因子：
6.1
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