Zhou Yang, Yang Daiwen
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore, 117543, Singapore.
J Biomol NMR. 2015 Feb;61(2):89-94. doi: 10.1007/s10858-014-9888-1. Epub 2014 Dec 4.
A new HSQC-based (13)Cα CEST pulse scheme is proposed, which is suitable for uniformly (13)C- or (13)C, (15)N-labeled samples in either water or heavy water. Except for Thr and Ser residues, the sensitivity of this scheme for uniformly labeled samples is similar to that of the previous scheme for selectively (13)Cα-labeled samples with 100% isotope enrichment. The experiment is demonstrated on an acyl carrier protein domain. Our (13)Cα CEST data reveal that the minor state of the acyl carrier protein has high helical propensity. The new scheme will facilitate structural characterization of invisible minor states.
提出了一种基于HSQC的新型(13)Cα CEST脉冲序列，该序列适用于水或重水中均匀(13)C标记或(13)C、(15)N双标记的样品。除苏氨酸和丝氨酸残基外，该序列对均匀标记样品的灵敏度与先前用于100%同位素富集的选择性(13)Cα标记样品的序列相似。在酰基载体蛋白结构域上进行了实验验证。我们的(13)Cα CEST数据表明，酰基载体蛋白的次要状态具有较高的螺旋倾向。新序列将有助于对不可见次要状态进行结构表征。
