Sensitivity-enhanced Four-dimensional Amide-amide Correlation NMR Experiments for Sequential Assignment of Proline-rich Disordered Proteins,Journal of the American Chemical Society

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Sensitivity-enhanced Four-dimensional Amide-amide Correlation NMR Experiments for Sequential Assignment of Proline-rich Disordered Proteins
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2018-02-28 , DOI: 10.1021/jacs.8b00215
Leo E. Wong ₁ , Joachim Maier ₁ , Jürgen Wienands ₂ , Stefan Becker ₁ , Christian Griesinger ₁

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Proline is prevalent in intrinsically disordered proteins (IDPs). NMR assignment of proline-rich IDPs is a challenge due to low dispersion of chemical shifts. We propose here new sensitivity-enhanced 4D NMR experiments that correlate two pairs of amide resonances that are either consecutive (NH i-1, NH i) or flanking a proline at position i-1 (NH i-2, NH i). The maximum 2-fold enhancement of sensitivity is achieved by employing two coherence order-selective (COS) transfers incorporated unconventionally into the pulse sequence. Each COS transfer confers an enhancement over amplitude-modulated transfer by a factor of √2 specifically when transverse relaxation is slow. The experiments connect amide resonances over a long fragment of sequence interspersed with proline. When this method was applied to the proline-rich region of B cell adaptor protein SLP-65 (pH 6.0) and α-synuclein (pH 7.4), which contain a total of 52 and 5 prolines, respectively, 99% and 92% of their nonprolyl amide resonances have been successfully assigned, demonstrating its robustness to address the assignment problem in large proline-rich IDPs.

中文翻译：

高灵敏度四维酰胺-酰胺相关核磁共振实验，用于序列指定富含脯氨酸的无序蛋白质

脯氨酸在本质上无序的蛋白质 (IDP) 中很普遍。由于化学位移的低分散性，富含脯氨酸的 IDP 的 NMR 分配是一个挑战。我们在这里提出了新的灵敏度增强 4D NMR 实验，该实验将两对酰胺共振关联起来，这些共振要么是连续的（NH i-1，NH i），要么是在 i-1 位（NH i-2，NH i）侧翼脯氨酸。通过采用两个非常规地合并到脉冲序列中的相干顺序选择性 (COS) 传输，最大程度地提高了 2 倍的灵敏度。每次 COS 传输都比调幅传输增强了 √2 倍，特别是当横向弛豫速度较慢时。实验将酰胺共振连接到散布有脯氨酸的长序列片段上。

更新日期：2018-02-28

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