Structural characterization and location of disulphide linkages of a potent vasodilatory peptide, recombinant maxadilan, by a multiple mass spectrometric approach.
Structural characterization and location of disulphide linkages of a potent vasodilatory peptide, recombinant maxadilan, by a multiple mass spectrometric approach.
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通过多重质谱方法对强效血管舒张肽重组 maxadilan 的二硫键进行结构表征和定位。
DOI：
10.1002/(sici)1097-0231(199604)10:6
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发表时间：
1996
影响因子：
2
通讯作者：
F. Kanda
中科院分区：
文献类型：
--
作者：
S. Yoshida;T. Takamatsu;S. Denda;M. Ohnuma;M. Tajima;E. Lerner;F. Kanda
采用快速原子轰击（FAB）和基质辅助激光解吸/电离（MALDI）的多重质谱策略已被用来确认序列，并定位重组maxadilan（r-maxadilan）（平均分子量7422.5 Da），一种有效的血管舒张肽从Lutzomyia longipalpis的二硫键。完整的r-马沙地兰，其减少的形式和其吡啶基乙基化的形式（p-马沙地兰）的MALDI测量表明，没有主要的翻译后修饰的四个半胱氨酸残基的存在。FAB和FAB-串联质谱法测量p-maxadilan的糜蛋白酶解酶足以绘制p-maxadilan的一级结构，尽管由于FAB中观察到的强烈抑制，MALDI的补充使用对于使用Asp-N消化的完整映射是必要的。通过比较r-maxadilan的胰蛋白酶解物还原前后的FAB质谱，实现Cys-5-Cys-9键的归属。由于含有Cys-18-Cys 55键的肽片段的分子量大于4000，因此MALDI测量对于该键的归属是必不可少的。结果充分支持了多重质谱策略在多肽和蛋白质结构表征中的价值。
A multiple mass spectrometric strategy using fast-atom bombardment (FAB) and matrix-assisted laser desorption/ionization (MALDI) has been used to confirm the sequence and to locate the disulfide linkages of recombinant maxadilan (r-maxadilan) (average molecular mass 7422.5 Da), a potent vasodilatory peptide from Lutzomyia longipalpis. MALDI measurements of intact r-maxadilan, its reduced form and its pyridylethylated form (p-maxadilan) indicated the presence of four Cys residues without major post-translational modifications. FAB and FAB-tandem mass spectrometry measurements of chymotryptic digests of p-maxadilan were sufficient to map the primary structure of p-maxadilan, though the complementary use of MALDI was necessary for complete mapping using Asp-N digestion due to a strong suppression observed in FAB. Assignment of the Cys-5-Cys-9 linkage was achieved by comparison of FAB mass spectra before and after reduction of tryptic digests of r-maxadilan. Since the molecular weight of the peptide fragment containing the Cys-18-Cys 55 linkage is more than 4000, MALDI measurement was indispensable for assignment of this linkage. The results fully support the value of the multiple mass spectrometric strategy in the structural characterization of peptides and proteins.