Lapeyre B, Amalric F, Ghaffari S H, Rao S V, Dumbar T S, Olson M O
J Biol Chem. 1986 Jul 15;261(20):9167-73.
By a combination of protein chemistry and recombinant DNA methods a glycine-rich region was found to be located near the carboxyl terminus of the nucleolar specific phosphoprotein, nucleolin, from Novikoff hepatoma (protein C23) and Chinese hamster ovary cells (100-kDa nucleolar protein). A sequence of 192 amino acid residues was derived from partial sequences of cyanogen bromide and N-bromosuccinimide fragments of protein C23 and deduced protein sequence from Chinese hamster ovary cell 100-kDa cDNA sequences. The 66 residues sequenced by protein methods were identical to the corresponding residues deduced by DNA sequencing. The multiple residues of NG,NG-dimethylarginine (DMA) contained in the nucleolin polypeptide were found to be limited to a segment of less than 10 kDa near the carboxyl-terminal end of the protein. This segment also contained internally repeated sequences (e.g. 7 copies of the sequence Gly-Gly-Arg-Gly-Gly were found) which were unrelated to sequences closer to the amino-terminal end. Most arginine residues in this region were surrounded by 2 or 3 glycine residues and were relatively close in sequence to phenylalanine residues.
通过蛋白质化学和重组DNA方法相结合,发现富含甘氨酸的区域位于来自诺维科夫肝癌细胞(蛋白质C23)和中国仓鼠卵巢细胞(100 kDa核仁蛋白)的核仁特异性磷蛋白核仁素的羧基末端附近。192个氨基酸残基的序列来自蛋白质C23的溴化氰和N-溴代琥珀酰亚胺片段的部分序列以及从中国仓鼠卵巢细胞100 kDa cDNA序列推导的蛋白质序列。通过蛋白质方法测序的66个残基与通过DNA测序推导的相应残基相同。发现核仁素多肽中含有的NG,NG-二甲基精氨酸(DMA)的多个残基仅限于蛋白质羧基末端附近小于10 kDa的片段。该片段还包含内部重复序列(例如,发现该序列Gly-Gly-Arg-Gly-Gly重复7次),这些序列与更靠近氨基末端的序列无关。该区域中的大多数精氨酸残基被2或3个甘氨酸残基包围,并且在序列上与苯丙氨酸残基相对接近。