Bond M D, Strydom D J
Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, Massachusetts 02115.
Biochemistry. 1989 Jul 11;28(14):6110-3. doi: 10.1021/bi00440a057.
The amino acid sequence and disulfide bridges of bovine plasma derived angiogenin were determined by sequencer analysis of the intact protein and fragments derived by enzymatic and chemical digestion. Bovine angiogenin is a single-chain protein of 125 amino acids; it contains six cysteines and has a calculated molecular weight of 14,595. In contrast to the human protein its amino terminus is unblocked. It has the following sequence: H2N-Ala1-Gln-Asp-Asp-Tyr-Arg-Tyr-Ile-His-Phe10-Leu-Thr-Gln-His-Tyr -Asp-Ala-Lys- Pro-Lys20-Gly-Arg-Asn-Asp-Glu-Tyr-Cys-Phe-Asn-Met30-Met-Lys- Asn-Arg-Arg-Leu-Thr - Arg-Pro-Cys40-Lys-Asp-Arg-Asn-Thr-Phe-Ile-His-Gly-Asn50-Lys- Asn-Asp-Ile-Lys-Ala - Ile-Cys-Glu-Asp60-Arg-Asn-Gly-Gln-Pro-Tyr-Arg-Gly-Asp-Leu70- Arg-Ile-Ser-Lys-Ser - Glu-Phe-Gln-Ile-Thr80-Ile-Cys-Lys-His-Lys-Gly-Ser-Ser-Arg90- Pro-Pro-Cys-Arg-Tyr - Gly-Ala-Thr-Glu-Asp100-Ser-Arg-Val-Ile-Val-Val-Gly-Cys-Glu-Asn1 10-Gly-Leu-Pro- Val-His-Phe-Asp-Glu-Ser-Phe120-Ile-Thr-Pro-Arg-His-OH. Disulfide bonds link Cys(27)-Cys(82), Cys(40)-Cys(93), and Cys(58)-Cys(108). Bovine angiogenin is 64% identical with human angiogenin; like the human protein, it is homologous to the pancreatic ribonucleases, with conservation of active site residues. Two regions, 6-22 and 65-75, are highly conserved between the angiogenins but are significantly different from those of the ribonucleases, suggesting a possible role in the molecules' biological activity.
通过对完整蛋白质以及酶解和化学消化产生的片段进行测序分析,确定了牛血浆来源血管生成素的氨基酸序列和二硫键。牛血管生成素是一种由125个氨基酸组成的单链蛋白质;它含有6个半胱氨酸,计算分子量为14,595。与人类蛋白质不同,其氨基末端未被封闭。它具有以下序列:H2N-Ala1-谷氨酰胺-天冬氨酸-天冬氨酸-酪氨酸-精氨酸-酪氨酸-异亮氨酸-组氨酸-苯丙氨酸10-亮氨酸-苏氨酸-谷氨酰胺-组氨酸-酪氨酸-天冬氨酸-丙氨酸-赖氨酸-脯氨酸-赖氨酸20-甘氨酸-精氨酸-天冬酰胺-天冬氨酸-谷氨酸-酪氨酸-半胱氨酸-苯丙氨酸-天冬酰胺-甲硫氨酸30-甲硫氨酸-赖氨酸-天冬酰胺-精氨酸-精氨酸-亮氨酸-苏氨酸-精氨酸-脯氨酸-半胱氨酸40-赖氨酸-天冬氨酸-精氨酸-天冬酰胺-苏氨酸-苯丙氨酸-异亮氨酸-组氨酸-甘氨酸-天冬酰胺50-赖氨酸-天冬酰胺-天冬氨酸-异亮氨酸-赖氨酸-丙氨酸-异亮氨酸-半胱氨酸-谷氨酸-天冬氨酸60-精氨酸-天冬酰胺-甘氨酸-谷氨酰胺-脯氨酸-酪氨酸-精氨酸-甘氨酸-天冬氨酸-亮氨酸70-精氨酸-异亮氨酸-丝氨酸-赖氨酸-丝氨酸-谷氨酸-苯丙氨酸-谷氨酰胺-异亮氨酸-苏氨酸80-异亮氨酸-半胱氨酸-赖氨酸-组氨酸-赖氨酸-甘氨酸-丝氨酸-丝氨酸-精氨酸90-脯氨酸-脯氨酸-半胱氨酸-精氨酸-酪氨酸-甘氨酸-丙氨酸-苏氨酸-谷氨酸-天冬氨酸100-丝氨酸-精氨酸-缬氨酸-异亮氨酸-缬氨酸-缬氨酸-甘氨酸-半胱氨酸-谷氨酸-天冬酰胺110-甘氨酸-亮氨酸-脯氨酸-缬氨酸-组氨酸-苯丙氨酸-天冬氨酸-谷氨酸-丝氨酸-苯丙氨酸120-异亮氨酸-苏氨酸-脯氨酸-精氨酸-组氨酸-OH。二硫键连接Cys(27)-Cys(82)、Cys(40)-Cys(93)和Cys(58)-Cys(108)。牛血管生成素与人类血管生成素的同源性为64%;与人类蛋白质一样,它与胰腺核糖核酸酶同源,活性位点残基保守。血管生成素之间的两个区域,6-22和65-75,高度保守,但与核糖核酸酶的区域明显不同,这表明它们可能在分子的生物学活性中发挥作用。