Zurmanová J, Maláčová D, Půta F, Novák P, Rícný J, Soukup T
Institute of Physiology AS CR, Prague, Czech Republic.
Physiol Res. 2007;56(5):659-662. doi: 10.33549/physiolres.931280.
We have separated 2b myosin heavy chain (MyHC) isoform from the rat extensor digitorum longus muscle by SDS-PAGE and analyzed it by two subsequent mass spectrometry techniques. After tryptic digestion, the obtained peptides were identified by Matrix-Assisted Laser Desorption/Ionisation reflectron Time of Flight mass spectrometry (MALDI-TOF MS) and sequenced by liquid chromatography tandem mass spectrometry (ESI LC/MS/MS). The analyzed peptides proportionally covered 30 % of the 2b MyHC isoform sequence. The results suggest that the primary structure is identical with the highest probability to a NCBI database record ref|NP_062198.1|, representing the last updated record of rat 2b isoform. Nonetheless, four peptides carrying amino acid substitution(s) in comparison with the NCBI database record were identified.
我们通过SDS-PAGE从大鼠趾长伸肌中分离出2b型肌球蛋白重链(MyHC)亚型,并通过两种后续的质谱技术对其进行分析。胰蛋白酶消化后,所得肽段通过基质辅助激光解吸/电离反射式飞行时间质谱(MALDI-TOF MS)进行鉴定,并通过液相色谱串联质谱(ESI LC/MS/MS)进行测序。分析的肽段按比例覆盖了2b型MyHC亚型序列的30%。结果表明,其一级结构与NCBI数据库记录ref|NP_062198.1|具有最高的一致性概率,该记录代表大鼠2b亚型的最新记录。尽管如此,还是鉴定出了四个与NCBI数据库记录相比存在氨基酸替换的肽段。