Selvakumar Ponniah, Lakshmikuttyamma Ashakumary, Pasha Mohammed Khysar, King Martin J, Olson Douglas J H, Mori Sumiko, Ross Andrew R S, Hayashi Kiyoshi, Dimmock Jonathan R, Sharma Rajendra K
Department of Pathology, College of Medicine and Research Unit, Saskatchewan Cancer Agency, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 4H4, Canada.
J Cell Biochem. 2004 Jun 1;92(3):573-8. doi: 10.1002/jcb.20085.
Many of viral and eukaryotic proteins are required for signal transduction and regulatory functions which undergo a lipid modification by the enzyme N-myristoyltransferase (NMT). In this study, we demonstrated that heat shock cognate protein 70 (HSC70) is homologous to NMT inhibitor protein (NIP71), which was discovered in our laboratory, based on MALDI-TOF mass spectrometric analysis. The purified bovine cytosolic HSC70 and particulate NIP71 produced a dose-dependent inhibition of human NMT having half maximal inhibitions of 235 and 230 nM, respectively. Further, Western blot analysis revealed that the purified particulate NIP71 and cytosolic HSC70 cross-reacted with both anti-NIP71 and anti-HSC70 antibodies. The results we obtained imply that molecular chaperones could be involved in the regulation of NMT in normal and cancerous cells. Further studies directed to revealing the role of HSC70 in the regulation of NMT may lead to the development of gene based therapies of colon cancer.
许多病毒蛋白和真核蛋白参与信号转导和调节功能,这些蛋白会被N-肉豆蔻酰转移酶(NMT)进行脂质修饰。在本研究中,基于基质辅助激光解吸电离飞行时间质谱分析,我们证明热休克同源蛋白70(HSC70)与我们实验室发现的NMT抑制蛋白(NIP71)同源。纯化的牛细胞质HSC70和颗粒状NIP71对人NMT产生剂量依赖性抑制,半数最大抑制浓度分别为235和230 nM。此外,蛋白质印迹分析表明纯化的颗粒状NIP71和细胞质HSC70与抗NIP71和抗HSC70抗体均发生交叉反应。我们获得的结果表明分子伴侣可能参与正常细胞和癌细胞中NMT的调节。进一步揭示HSC70在NMT调节中作用的研究可能会促成基于基因的结肠癌治疗方法的开发。