Garoff H, Riedel H, Lehrach H
Nucleic Acids Res. 1982 Jan 22;10(2):675-87. doi: 10.1093/nar/10.2.675.
We describe a peptide sequencing procedure which can be used to verify an amino acid sequence which is derived from a nucleotide sequence. One first labels the protein with a 3H- and a 14C-labelled amino acid and then cleaves the protein into a set of peptides using a cleavage reaction specific for a particular amino acid residue. Finally one performs Edman degradations on the whole mixture of peptides. The released amino acids reflect the combined aminoterminal amino acid sequences of all the peptides that have been formed by the cleavage reaction. The data can therefore be used to check a deduced sequence simultaneously at several regions of the polypeptide chain. We have applied this sequencing procedure to verify the amino acid sequence deduced from the 26S RNA of Semliki Forest virus.
我们描述了一种肽测序方法,该方法可用于验证从核苷酸序列推导而来的氨基酸序列。首先用3H和14C标记的氨基酸对蛋白质进行标记,然后使用针对特定氨基酸残基的裂解反应将蛋白质切割成一组肽段。最后,对整个肽段混合物进行埃德曼降解。释放出的氨基酸反映了由裂解反应形成的所有肽段的组合氨基末端氨基酸序列。因此,这些数据可用于同时在多肽链的多个区域检查推导的序列。我们已应用此测序方法来验证从塞姆利基森林病毒26S RNA推导的氨基酸序列。