Sequential 1H NMR assignments and secondary structure of hen egg white lysozyme in solution.
Sequential 1H NMR assignments and secondary structure of hen egg white lysozyme in solution.
复制标题
溶液中鸡蛋清溶菌酶的连续 1H NMR 归属和二级结构。
作者：
Christina Redfield;Christopher M. Dobson
通过序列特异性方法获得了蛋清溶菌酶129个残基中121个的1H NMR谱。采用相敏二维（2-D）相关光谱和单、双中继相干转移光谱对自旋体系进行了鉴定。对于关键类型的氨基酸残基，特别是丙氨酸、苏氨酸、缬氨酸和甘氨酸，完整的自旋体系已经确定。对于其他残基，发现自旋体系的一个不太完整的定义足以满足顺序分配的目的。序列特异性定位是通过相敏二维核Overhauser增强光谱（NOESY）实现的。利用溶菌酶中广泛的氢交换率是克服光谱重叠问题的有效方法。序列分配由21个肽段建立，长度从2到13个残基不等。NOESY光谱还用于提供有关溶液中蛋白质二级结构的信息。从NOESY数据中鉴定出3个螺旋区和2个β片区；这些区域与母鸡溶菌酶的x射线结构相同。缓慢交换的酰胺通常与x射线结构中发现的氢键有关；但是，也发现了这一总趋势的一些例外情况。本文的结果表明，从蛋白质的二维核磁共振光谱中可以获得非常详细的信息，这些信息比以前的研究要大得多。
Assignments for 1H NMR resonances of 121 of the 129 residues of hen egg white lysozyme have been obtained by sequence-specific methods. Spin systems were identified with phase-sensitive two-dimensional (2-D) correlated spectroscopy and single and double relayed coherence transfer spectroscopy. For key types of amino acid residues, particularly alanine, threonine, valine, and glycine, complete spin systems were identified. For other residues a less complete definition of the spin system was found to be adequate for the purpose of sequential assignment. Sequence-specific assignments were achieved by phase-sensitive 2-D nuclear Overhauser enhancement spectroscopy (NOESY). Exploitation of the wide range of hydrogen exchange rates found in lysozyme was a useful approach to overcoming the problem of spectral overlap. The sequential assignment was built up from 21 peptide segments ranging in length from 2 to 13 residues. The NOESY spectra were also used to provide information about the secondary structure of the protein in solution. Three helical regions and two regions of beta-sheet were identified from the NOESY data; these regions are identical with those found in the X-ray structure of hen lysozyme. Slowly exchanging amides are generally correlated with hydrogen bonding identified in the X-ray structure; a number of exceptions to this general trend were, however, found. The results presented in this paper indicate that highly detailed information can be obtained from 2-D NMR spectra of a protein that is significantly larger than those studied previously.
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