Bures E J, Nika H, Chow D T, Morrison H D, Hess D, Aebersold R
Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
Anal Biochem. 1995 Jan 1;224(1):364-72. doi: 10.1006/abio.1995.1052.
We report the synthesis and structural characterization of the novel Edman-type protein-sequencing reagent 4-(3-pyridinylmethylaminocarboxypropyl) phenyl isothiocyanate. A panel of thiohydantoins prepared from this reagent were found stable during liquid chromatography-electrospray mass spectrometry and were detectable at the low femtomole sensitivity level. Furthermore, the signal detected for these compounds in the mass spectrometer was linear from the low femtomole to the low picomole range. The derivatives showed uv absorbance spectra comparable to their phenylthiohydantoin counterparts. The extinction coefficient for the 4-(3-pyridinylmethylaminocarboxypropyl) phenyl thiohydantoin tyrosine was determined by adsorptive sequence analysis of a synthetic pentapeptide featuring an N-terminal 125I-labeled tyrosine. The sequence data suggest that the reagent will be useful for extended sequence analysis of proteins and peptides using commercially available gas-liquid-phase sequencers.
我们报道了新型埃德曼型蛋白质测序试剂4-(3-吡啶基甲基氨基羧丙基)苯基异硫氰酸酯的合成及结构表征。由该试剂制备的一组乙内酰苯硫脲在液相色谱-电喷雾质谱分析过程中被发现是稳定的，并且在低飞摩尔灵敏度水平下可被检测到。此外，在质谱仪中检测到的这些化合物的信号在低飞摩尔到低皮摩尔范围内呈线性。这些衍生物的紫外吸收光谱与其苯硫代乙内酰脲对应物相当。4-(3-吡啶基甲基氨基羧丙基)苯基硫代乙内酰脲酪氨酸的消光系数通过对一个以N端125I标记酪氨酸为特征的合成五肽进行吸附序列分析来确定。序列数据表明，该试剂将有助于使用市售气液相测序仪对蛋白质和肽进行扩展序列分析。
