Assignment of selected hyperfine proton NMR resonances in the met forms of Glycera dibranchiata monomer hemoglobins and comparisons with sperm whale metmyoglobin.
Assignment of selected hyperfine proton NMR resonances in the met forms of Glycera dibranchiata monomer hemoglobins and comparisons with sperm whale metmyoglobin.
复制标题
选定的超精细质子核磁共振共振在双臂甘油单体血红蛋白的met形式中的分配以及与抹香鲸met肌红蛋白的比较。
DOI：
10.1021/bi00408a059
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Smith,KM
中科院分区：
文献类型：
--
作者：
Constantinidis,I;Satterlee,JD;Pandey,RK;Leung,HK;Smith,KM
加州大学化学系，戴维斯，加州，95616接收于1987年8月27日;修订后的Mandarin pt接收于1987年12月10日摘要：这项工作表明，我们制备的所有三种主要的二鳃甘油单体血红蛋白的纯度很高，并详细说明了亚铁（水？）蛋白质形式。通过用选择性氘代血红素重组每个组分的脱辅基蛋白进行分配。结果表明，即使单个组分制剂基本上由单一蛋白质组成，质子NMR光谱也表明光谱异质性。证据提出的主要和次要的蛋白质形式，存在于每个组件的解决方案的识别和分类。最后，与以前的结果相比，详细分析了每个组分的主要和次要形式的质子超精细移位模式，与高铁肌红蛋白的主要和次要形式相比，得出的结论是，来自每个物种的蛋白质的对应形式具有惊人相似的血红素-珠蛋白接触，并显示几乎相同的血红素电子结构和配位数。海洋环节动物二鳃甘油的红细胞最初显示含有三种主要的单体血红蛋白（组分II、III和IV），它们可以容易地分离（Kandler和Satterlee，1983; Kandler等人，1984年）。后来的工作证实了我们最初的结果，并产生了关于这些蛋白质的光谱独特性的补充信息（Cooke & Wright，1985 a，B; Constantinavian & Satterlee，1987）。对这些蛋白质的主要兴趣起源于晶体学（Padlan & Love，1974）和测序（Imamura等人，1972）均表明在一个单体血红蛋白中缺少远端组氨酸（E-7）。在它的位置上发现了亮氨酸。这种替代的影响似乎立即被认识到。其中一个G. dibranchiata血红蛋白与肌红蛋白的同源性（Padlan & Love，1974）和它们的可比序列同源性（Imamura等人，1972; Satterlee，1984）使这些蛋白质成为研究血红素-珠蛋白结构-功能关系的工作者的直接兴趣（Escheronds等人，1976年; Eschonds & Forster，1972年; Eschonds，1971年; Weber等人，1977; Parkhurst等人，1980年）。其原因是，与具有通常为极性或带电氨基酸的特殊E-7取代的肌红蛋白或血红蛋白不同（Romero-Herrera等人，1981; Dene等人，1980; Huber等人，1971; Wollmer等人，1971），G. dibranchiata取代是用具有完全烃侧链的氨基酸（亮氨酸）取代远端组氨酸的取代。鉴于对这些独特蛋白质的兴趣，我们的目标是f的生物化学、光谱和动力学表征。这项工作得到了美国国立卫生研究院的支持，JDS获得了赠款DK 30912和HL 01758（研究职业发展奖），KMS获得了HL 22252。
Department of Chemistry, University of California, Davis, California 95616 Received August 27, 1987; Revised Manuscript Received December 10, 1987 abstract: This work indicates a high degree of purity for our preparations of all three of the primary Glycera dibranchiata monomer hemoglobins and details assignments of the heme methyl and vinyl protons in the hyperfine shift region of the ferric (aquo?) protein forms. The assignments were carried out by reconstituting the apoproteins of each component with selectively deuteriated hemes. The results indicate that even though the individual component preparations consist of essentially a single protein, the proton NMR spectra indicate spectroscopic heterogeneity. Evidence is presented for identification and classification of major and minor protein forms that are present in solutions of each component. Finally, in contrast to previous results, a detailed analysis of the proton hyperfineshift patterns of the major and minor forms of each component, in comparison to the major and minor forms of metmyoglobin, leads to the conclusion that thecorresponding forms of the proteins from each species have strikingly similar heme-globin contacts and display nearly identical heme electronic structures and coordination numbers. e erythrocytes of the marine annelid Glycera dibranchiata were originally shown to contain threemajor monomeric hemoglobins (components II, III, and IV) that could be easily isolated (Kandler& Satterlee, 1983; Kandler et al., 1984). Later work confirmed our original results and produced ad-ditional information aboutthe spectroscopic uniqueness of these proteins (Cooke & Wright, 1985a, b; Constantinidis & Satterlee, 1987). The principal interest in these proteins or-iginates from early work in which crystallography (Padlan & Love, 1974) and sequencing (Imamura et al., 1972) both indicated that the distal histidine (E-7) was missing in one of the monomer hemoglobins. In its place leucine was found. The implications of this substitution seem to have been immediately realized. The overall three-dimensional structural resemblance of one of the G. dibranchiata hemoglobins to myoglobins (Padlan & Love, 1974) and their comparable sequence homologies (Imamura et al., 1972; Satterlee, 1984) made these proteins of immediate interest to workers studying heme-globin structure-function relationships (Seamonds et al., 1976; Seamonds & Forster, 1972; Seamonds, 1971; Weber et al., 1977; Parkhurst et al., 1980). The reason for this is that unlike myoglobins or hemoglobins that possess exceptional E-7 substitutions which are generally polar or charged amino acids (Romero-Herrera et al., 1981; Dene et al., 1980; Huber et al., 1971; Wollmer et al., 1971) the G. dibranchiata sub-stitution is one that replaces the distal histidine by an amino acid with a completely hydrocarbon side chain (leucine). Given the interest in these unique proteins, our goal has been biochemical, spectroscopic, and kinetics characterization of f This work was supported by the National Institutes of Health through Grants DK30912 and HL01758 (Research Career Development Award) to JDS and HL22252 to KMS Additional support came from the Alfred P. Sloan Foundation through a fellowship awardedto JDS