Sen R, Sharp R R
Biochim Biophys Acta. 1980 Jul 3;630(3):447-58. doi: 10.1016/0304-4165(80)90294-9.
Carbon-13 NMR spectra of the reconcentrated chromaffin granule lysate have been obtained at 50 MHz and 62.9 MHz. The spectrum contains a number of assignable resonances in addition to those of the main soluble components (catecholamines, adenine nucleotides and chromogranin). Guanine and uridine nucleotides are present at levels of 0.13 and 0.08 mol/mol adenine nucleotides, respectively. Concentrations of cytidine nucleotides and NAD+ are below the detection limit (0.02 mol/mol adenine nucleotides). An unidentified low molecular weight species, thought to be an adenine-containing oligonucleotide, is also present. Ascorbic acid was observed at a concentration of 0.14 mol/mol adenine nucleotides, but both dopamine and dehydroascorbic acid were below the detection limit. Protein resonances agree well with the reported amino acid composition of chromogranin A, with the exception of tryptophan and glutamine which have not previously been measured. The concentrations of these residues are estimated to be 12 +/- 3 and 39 +/- 5 residues per 77 000 dalton unit of chromogranin A. Substantial intensity due to unsaturated fatty acid side-chains in solubilized lipid is seen in the olefinic carbon region and in the methylene region, suggesting the presence of lipoprotein. Unassigned carbohydrate resonances are also present, but are largely obscured by sucrose in the isolation medium.
已在50兆赫和62.9兆赫下获得了重新浓缩的嗜铬粒蛋白颗粒裂解物的碳-13核磁共振谱。该谱除了主要可溶性成分(儿茶酚胺、腺嘌呤核苷酸和嗜铬粒蛋白)的共振峰外,还包含许多可归属的共振峰。鸟嘌呤和尿嘧啶核苷酸的含量分别为0.13摩尔/摩尔腺嘌呤核苷酸和0.08摩尔/摩尔腺嘌呤核苷酸。胞嘧啶核苷酸和NAD+的浓度低于检测限(0.02摩尔/摩尔腺嘌呤核苷酸)。还存在一种未鉴定的低分子量物质,被认为是一种含腺嘌呤的寡核苷酸。观察到抗坏血酸的浓度为0.14摩尔/摩尔腺嘌呤核苷酸,但多巴胺和脱氢抗坏血酸均低于检测限。蛋白质共振峰与报道的嗜铬粒蛋白A的氨基酸组成非常吻合,但色氨酸和谷氨酰胺除外,此前尚未对它们进行测量。估计这些残基的浓度为每77000道尔顿嗜铬粒蛋白A单位有12±3个色氨酸残基和39±5个谷氨酰胺残基。在烯烃碳区域和亚甲基区域可以看到由于溶解脂质中不饱和脂肪酸侧链产生的大量信号强度,表明存在脂蛋白。也存在未归属的碳水化合物共振峰,但在很大程度上被分离介质中的蔗糖所掩盖。