LeBoeuf R C, Miller C, Shively J E, Schumaker V N, Balla M A, Lusis A J
FEBS Lett. 1984 May 7;170(1):105-8. doi: 10.1016/0014-5793(84)81378-2.
A successful approach has been developed for the sequencing of apolipoprotein B based upon the procedure of Cleveland et al. [(1977) J. Biol. Chem. 252, 1102-1106] involving limited proteolysis in the presence of sodium dodecyl sulfate. Staphylococcus aureus protease was employed to produce large peptides which were isolated in relatively pure form by preparative gel electrophoresis. Two peptides were partially sequenced using spinning-cup microsequencing techniques. The sequences are: Peptide R2-5, -Ala-Leu-Val-Gly-Ile-Asn- Gly-Glu-Ala-Asn-Leu-Asp-Phe-Leu-Asn-Ile-Pro-Leu-Arg-Ile-Pro-Pro- Met-Arg-(Arg)-; Peptide R3-1, -Leu-Val-Ala-Lys-Pro-Ser-Val-Ser-Val-Glu- Phe-Val-Thr-Asn-Met-Gly-Ile-Ile-Pro-Lys-Phe-Ala-Arg-. Several stretches of residues suitable for the construction of oligonucleotide probes have been identified.
基于克利夫兰等人[（1977年）《生物化学杂志》252卷，1102 - 1106页]的方法，已经开发出一种成功的载脂蛋白B测序方法，该方法涉及在十二烷基硫酸钠存在下进行有限的蛋白酶解。使用金黄色葡萄球菌蛋白酶产生大的肽段，这些肽段通过制备性凝胶电泳以相对纯的形式分离出来。使用旋转杯微测序技术对两个肽段进行了部分测序。序列如下：肽段R2 - 5，-丙氨酸-亮氨酸-缬氨酸-甘氨酸-异亮氨酸-天冬酰胺-甘氨酸-谷氨酸-丙氨酸-天冬酰胺-亮氨酸-天冬氨酸-苯丙氨酸-亮氨酸-天冬酰胺-异亮氨酸-脯氨酸-亮氨酸-精氨酸-异亮氨酸-脯氨酸-脯氨酸-甲硫氨酸-精氨酸-（精氨酸）-；肽段R3 - 1，-亮氨酸-缬氨酸-丙氨酸-赖氨酸-脯氨酸-丝氨酸-缬氨酸-丝氨酸-缬氨酸-谷氨酸-苯丙氨酸-缬氨酸-苏氨酸-天冬酰胺-甲硫氨酸-甘氨酸-异亮氨酸-异亮氨酸-脯氨酸-赖氨酸-苯丙氨酸-丙氨酸-精氨酸-。已经鉴定出几段适合构建寡核苷酸探针的残基。
