Lee J, Lentz B R
Department of Biochemistry and Biophysics, University of North Carolina, School of Medicine, Chapel Hill, NC 27599-7260, USA.
Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9274-9. doi: 10.1073/pnas.95.16.9274.
Activation energies for the individual steps of secretory and viral fusion are reported to be large [Oberhauser, A. F., Monck, J. R. & Fernandez, J. M. (1992) Biophys. J. 61, 800-809; Clague, M. J., Schoch, C., Zech, L. & Blumenthal, R. (1990) Biochemistry 29, 1303-1308]. Understanding the cause for these large activation energies is crucial to defining the mechanisms of these two types of biological membrane fusion. We showed recently that the fusion of protein-free model lipid bilayers mimics the sequence of steps observed during secretory and viral fusion, suggesting that these processes may involve common lipid, rather than protein, rearrangements. To test for this possibility, we determined the activation energies for the three steps that we were able to distinguish as contributing to the fusion of protein-free model lipid bilayers. Activation energies for lipid rearrangements associated with formation of the reversible first intermediate, with conversion of this to a semi-stable second intermediate, and with irreversible fusion pore formation were 37 kcal/mol, 27 kcal/mol, and 22 kcal/mol, respectively. The first and last of these were comparable to the activation energies observed for membrane lipid exchange (42 kcal/mol) during viral fusion and for the rate of fusion pore opening during secretory granule release (23 kcal/mol). This striking similarity suggests strongly that the basic molecular processes involved in secretory and viral fusion involve a set of lipid molecule rearrangements that also are involved in model membrane fusion.
据报道,分泌性融合和病毒融合各个步骤的活化能都很高[奥伯豪泽,A. F.,蒙克,J. R. & 费尔南德斯,J. M.(1992年)《生物物理学杂志》61卷,800 - 809页;克拉格,M. J.,朔赫,C.,泽赫,L. & 布卢门撒尔,R.(1990年)《生物化学》29卷,1303 - 1308页]。了解这些高活化能的成因对于界定这两种生物膜融合机制至关重要。我们最近表明,无蛋白模型脂质双层的融合模拟了分泌性融合和病毒融合过程中观察到的步骤顺序,这表明这些过程可能涉及共同的脂质重排,而非蛋白质重排。为了验证这种可能性,我们确定了我们能够区分的、对无蛋白模型脂质双层融合有贡献的三个步骤的活化能。与可逆的第一个中间体形成、该中间体转化为半稳定的第二个中间体以及不可逆的融合孔形成相关的脂质重排的活化能分别为37千卡/摩尔、27千卡/摩尔和22千卡/摩尔。其中第一个和最后一个与病毒融合过程中膜脂质交换的活化能(42千卡/摩尔)以及分泌颗粒释放过程中融合孔开放速率(23千卡/摩尔)相当。这种显著的相似性强烈表明,分泌性融合和病毒融合所涉及的基本分子过程涉及一组脂质分子重排,这些重排也参与了模型膜融合。