Fullmer C S, Wasserman R H
J Biol Chem. 1981 Jun 10;256(11):5669-74.
The complete amino acid sequence of the vitamin D-dependent bovine intestinal calcium-binding protein (minor A component) has been determined: Lys-Ser-Pro-Glu-Glu-Leu-Lys-Gly-Ile-Phe-Glu-Lys-Tyr-Ala-Ala-Lys-Glu-Gly-Asp-Pro-Asn-Gln-Leu-Ser-Lys-Glu-Glu-Leu-Lys-Leu-Leu-Leu-Gln-Thr-Glu-Phe-Pro-Ser-Leu-Le u-Lys-Gly-Pro-Ser-Thr-Leu-Asp-Glu-Leu-Phe-Glu-Glu-Leu-Asp-Lys-Asn-Gly-Asp-Gly-Glu-Val-Ser-Phe-Glu-Glu-Phe-Gln-Val-Leu-Val-Lys-Lys-Ile-Ser-Gln-OH. It is a 75-residue protein (computed Mr = 8501), contains a single Tyr, and is devoid of Cys, Met, Trp, His, and Arg. The bulk of the sequence was determined by automated sequencing of: (i) the intact protein for 20 cycles; (ii) a large N-bromosuccinimide peptide for 37 cycles; (iii) a tryptic peptide (29 cycles), isolated by high performance liquid chromatography. Also described is a highly sensitive and rapid procedure for peptide mapping by high performance liquid chromatography.
已确定维生素D依赖性牛肠钙结合蛋白(次要A组分)的完整氨基酸序列:赖氨酸-丝氨酸-脯氨酸-谷氨酸-谷氨酸-亮氨酸-赖氨酸-甘氨酸-异亮氨酸-苯丙氨酸-谷氨酸-赖氨酸-酪氨酸-丙氨酸-丙氨酸-赖氨酸-谷氨酸-甘氨酸-天冬氨酸-脯氨酸-天冬酰胺-谷氨酰胺-亮氨酸-丝氨酸-赖氨酸-谷氨酸-谷氨酸-亮氨酸-赖氨酸-亮氨酸-亮氨酸-亮氨酸-谷氨酰胺-苏氨酸-谷氨酸-苯丙氨酸-脯氨酸-丝氨酸-亮氨酸-亮氨酸-赖氨酸-甘氨酸-脯氨酸-丝氨酸-苏氨酸-亮氨酸-天冬氨酸-谷氨酸-亮氨酸-苯丙氨酸-谷氨酸-谷氨酸-亮氨酸-天冬氨酸-赖氨酸-天冬酰胺-甘氨酸-天冬氨酸-甘氨酸-谷氨酸-缬氨酸-丝氨酸-苯丙氨酸-谷氨酸-谷氨酸-苯丙氨酸-谷氨酰胺-缬氨酸-亮氨酸-缬氨酸-赖氨酸-赖氨酸-异亮氨酸-丝氨酸-谷氨酰胺-OH。它是一种含75个残基的蛋白质(计算分子量=8501),含有一个酪氨酸,不含半胱氨酸、甲硫氨酸、色氨酸、组氨酸和精氨酸。序列的大部分是通过以下自动测序确定的:(i)完整蛋白质测序20个循环;(ii)一个大的N-溴代琥珀酰亚胺肽测序37个循环;(iii)一个通过高效液相色谱分离的胰蛋白酶肽(测序29个循环)。还描述了一种通过高效液相色谱进行肽图谱分析的高度灵敏且快速的方法。