CAJ | 학술논문

小批量制备质谱纯鲨鱼肝铁蛋白(Liver Ferritin of Sphyrna zygaena, SZLF).在弱酸介质(pH 1.0)中,天然电泳结果显示,SZLF蛋白质亚基20 min后开始解离.选用透射电子显微镜跟踪SZLF亚基解离与重组装全过程和蛋白壳与铁核尺寸变化, 发现SZLF在亚基酸解离过程中,随着pH值的降低,铁核和蛋白壳的尺寸呈现相同的变化趋势,这种变化趋势可能与铁核内层铁的释放和蛋白壳的解离与去折叠有关.SZLF蛋白壳的重组装过程则是一个快速过程,并且是由松散熔球态向紧密态转变的过程.SZLF由单类型亚基组成,而马脾铁蛋白(Horse Spleen Ferritin , HSF)由H和L两种亚基类型组成.在基质pH 3.0条件和激光辅助下,混合HSF和SZLF仍然可释放各自的亚基且形成准亚基离子,供基质辅助激光解析电离飞行时间质谱分析,说明此时SZLF的亚基间相互作用强度减弱但并没有去折叠.TEM技术在铁蛋白解离和重组装过程中的应用,为进一步研究铁蛋白纳米包装的过程和机理提供新颖的、可行的和更加直观的研究手段.
소비량제비질보순사어간철단백(Liver Ferritin of Sphyrna zygaena, SZLF).재약산개질(pH 1.0)중,천연전영결과현시,SZLF단백질아기20 min후개시해리.선용투사전자현미경근종SZLF아기해리여중조장전과정화단백각여철핵척촌변화, 발현SZLF재아기산해리과정중,수착pH치적강저,철핵화단백각적척촌정현상동적변화추세,저충변화추세가능여철핵내층철적석방화단백각적해리여거절첩유관.SZLF단백각적중조장과정칙시일개쾌속과정,병차시유송산용구태향긴밀태전변적과정.SZLF유단류형아기조성,이마비철단백(Horse Spleen Ferritin , HSF)유H화L량충아기류형조성.재기질pH 3.0조건화격광보조하,혼합HSF화SZLF잉연가석방각자적아기차형성준아기리자,공기질보조격광해석전리비행시간질보분석,설명차시SZLF적아기간상호작용강도감약단병몰유거절첩.TEM기술재철단백해리화중조장과정중적응용,위진일보연구철단백납미포장적과정화궤리제공신영적、가행적화경가직관적연구수단.
Liver ferritin of Sphyrna zygaena(SZLF) with purity of mass spectrum was prepared in batch. Under) the condition of acidifying medium at pH 1.0, PAGE showed that SZLF subunits treated for 20 min began) to dissociate. A whole process of subunit dissociation and recombination was monitored by transmission electron microscopy(TEM). In addition, the changes of size of both protein shell and iron core were also determined) by TEM directly. It was found that in the acid dissociation process of SZLF subunits, the size of iron) core and protein shell showed the same trend of change, which might be related to not only the iron release) of inner iron core but the dissociation and unfolding of the protein shell. The passway of SZLF recombination is a fast step, which is a conversion process from incompact moltenglobule to compact ferritin. Under the assistant of matrix acidity pH 3.0 and laser, SZLF mixed with horse spleen ferritin(HSF) still has capacity to release) its subunits to form subunit ions for mass analysis by a MALDI-TOF mass spectrometer, which indicates that the interaction intensity between the subunits was weaken but they were not unfolded under this pH condition. TEM technology can be applied in studying both dissociation and recombination in ferritin subunits.