Thiede B, von Janta-Lipinski M
Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Rapid Commun Mass Spectrom. 1998;12(23):1889-94. doi: 10.1002/(SICI)1097-0231(19981215)12:23<1889::AID-RCM411>3.0.CO;2-0.
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used to explore noncovalent interactions between different peptides and ribose nucleic acids (RNAs). One RNA was mixed together with two or more peptides or vice versa to compare the different effects of the molecules for noncovalent complex formation. The matrix 2,4,6-trihydroxyacetophenone was considered optimal for these studies due to the fact that peptides and RNA showed roughly the same peak intensities, in negative ion mode, as well as RNA-peptide complexes being detected. The formation of the noncovalent RNA-peptide complexes showed a correlation with the number of the basic amino acids arginine, lysine and histidine. The strongest influence of these amino acids for complex formation was obtained with arginine. Although different RNA molecules were used with different compositions and secondary structures, no specific effects to complex formation was observed. The comparison of noncovalent complexes with covalent RNA-peptide complexes, which were obtained from ribosomal subunits after cross-linking and enzymatic cleavages, showed that the specific RNA-protein interactions are dependent on the three-dimensional structure of the ribosome and its components. The results of this report indicate that MALDI-MS may be useful for the study of noncovalent interactions, in particular for peptides and RNA.
基质辅助激光解吸/电离质谱法（MALDI-MS）被用于探究不同肽与核糖核酸（RNA）之间的非共价相互作用。将一种RNA与两种或更多种肽混合在一起，反之亦然，以比较这些分子对非共价复合物形成的不同影响。基质2,4,6-三羟基苯乙酮被认为是这些研究的最佳选择，因为在负离子模式下，肽和RNA显示出大致相同的峰强度，并且还能检测到RNA-肽复合物。非共价RNA-肽复合物的形成与碱性氨基酸精氨酸、赖氨酸和组氨酸的数量相关。这些氨基酸对复合物形成的最强影响是由精氨酸产生的。尽管使用了具有不同组成和二级结构的不同RNA分子，但未观察到对复合物形成的特定影响。将非共价复合物与通过交联和酶切从核糖体亚基获得的共价RNA-肽复合物进行比较，结果表明特定的RNA-蛋白质相互作用取决于核糖体及其组分的三维结构。本报告的结果表明，MALDI-MS可能有助于研究非共价相互作用，特别是对于肽和RNA。
