Lucas F, Barber M, Wolstenholme W A, Geddes A J, Graham G N, Morris H R
Biochem J. 1969 Oct;114(4):695-702. doi: 10.1042/bj1140695.
Several peptides were isolated from the protein silk fibroin of Bombyx mori by means of ion-exchange chromatography of a chymotryptic digest. The sequences of three of the peptides, Gly-Ala-Gly-Tyr, Gly-Val-Gly-Tyr and Gly-Ala-Gly-Ala-Gly-Ala-Gly-Tyr, were known from previous chemical work, but the sequence of the fourth, Gly-Ala-Gly-Val-Gly-Ala-Gly-Tyr, was previously only partially known. The necessary volatility for mass-spectrometric examination of the peptides was achieved by permethylation of the N-acetyl-peptide methyl ester derivatives. From the mass spectra it was possible to confirm the known sequences and to establish that of the partially known one. In one instance it was possible to deduce from the same mass spectrum the sequence of a main peptide component and that of a small amount of contaminating peptide. These results demonstrate for the first time the use of mass spectrometry in the determination of the amino acid sequences in peptides from a protein hydrolysate.
通过胰凝乳蛋白酶消化产物的离子交换色谱法，从家蚕的丝心蛋白中分离出了几种肽。其中三种肽的序列，即甘氨酸-丙氨酸-甘氨酸-酪氨酸、甘氨酸-缬氨酸-甘氨酸-酪氨酸和甘氨酸-丙氨酸-甘氨酸-丙氨酸-甘氨酸-丙氨酸-甘氨酸-酪氨酸，已在之前的化学研究中知晓，但第四种肽，即甘氨酸-丙氨酸-甘氨酸-缬氨酸-甘氨酸-丙氨酸-甘氨酸-酪氨酸，此前仅部分已知。通过对N-乙酰肽甲酯衍生物进行全甲基化，实现了肽进行质谱分析所需的挥发性。从质谱图中可以确认已知序列，并确定部分已知序列的肽的序列。在一个实例中，能够从同一张质谱图中推断出主要肽成分的序列以及少量污染肽的序列。这些结果首次证明了质谱法在测定蛋白质水解产物中肽的氨基酸序列方面的应用。
