Ovádi Judit, Saks Valdur
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Budapest, Hungary.
Mol Cell Biochem. 2004 Jan-Feb;256-257(1-2):5-12. doi: 10.1023/b:mcbi.0000009855.14648.2c.
The history of the development of the ideas and research of organized metabolic systems during last three decades is shortly reviewed. The cell cytoplasm is crowded with solutes, soluble macromolecules such as enzymes, nucleic acids, structural proteins and membranes. The high protein density within the large compartments of the cells predominantly determines the major characteristics of cellular environment such as viscosity, diffusion and inhomogeneity. The fact that the solvent viscosity of cytoplasm is not substantially different from the water is explained by intracellular structural heterogeneity: the intrinsic macromolecular density is relatively low within the interstitial voids in the cell because many soluble enzymes are apparently integral parts of the insoluble cytomatrix and are not distributed homogeneously. The molecular crowding and sieving restrict the mobility of very large solutes, binding severely restrict the mobility of smaller solutes. One of consequence of molecular crowding and hindered diffusion is the need to compartmentalize metabolic pathway to overcome diffusive barriers. Although the movement of small molecules is slowed down in the cytoplasm, the metabolism can successfully proceed and even be facilitated by metabolite channeling which directly transfers the intermediate from one enzyme to an adjacent enzyme without the need of free aqueous-phase diffusion. The enhanced probability for intermediates to be transferred from one active site to the other by sequential enzymes requires stable or transient interactions of the relevant enzymes, which associate physically in non-dissociable, static multienzyme complexes--metabolones, particles containing enzymes of a part or whole metabolic systems. Therefore, within the living cell the metabolism depends on the structural organization of enzymes forming microcompartments. Since cells contain many compartments and microenvironments, the measurement of the concentration of metabolites in whole cells or tissues gives an average cellular concentration and not that which is actually sensed by the active site of a specific enzyme. Thus, the microcompartmentation could provide a mechanism which can control metabolic pathways. Independently and in parallel to the developments described above, the ideas of compartmentation came into existence from the necessity to explain important physiological phenomena, in particular in heart research and in cardiac electrophysiology. These phenomena demonstrated the physiological importance of the biophysical and biochemical mechanisms described in this review.
本文简要回顾了过去三十年中有关有组织代谢系统的思想和研究的发展历程。细胞质中充满了溶质、可溶性大分子,如酶、核酸、结构蛋白和膜。细胞大隔室内的高蛋白密度主要决定了细胞环境的主要特征,如粘度、扩散和不均匀性。细胞质的溶剂粘度与水没有实质性差异这一事实,可通过细胞内结构的异质性来解释:由于许多可溶性酶显然是不溶性细胞基质的组成部分且分布不均一,细胞间质空隙内的固有大分子密度相对较低。分子拥挤和筛分限制了非常大的溶质的移动性,而结合则严重限制了较小溶质的移动性。分子拥挤和扩散受阻的一个后果是需要将代谢途径分隔开来以克服扩散障碍。尽管小分子在细胞质中的移动速度减慢,但代谢仍可成功进行,甚至可通过代谢物通道化得到促进,代谢物通道化可直接将中间体从一种酶转移到相邻酶,而无需自由水相扩散。中间体通过顺序酶从一个活性位点转移到另一个活性位点的概率增加,这需要相关酶之间稳定或短暂的相互作用,这些酶在不可解离的静态多酶复合物——代谢体(包含部分或整个代谢系统酶的颗粒)中物理结合。因此,在活细胞内,代谢依赖于形成微区室的酶的结构组织。由于细胞包含许多区室和微环境,测量全细胞或组织中代谢物的浓度得到的是平均细胞浓度,而不是特定酶的活性位点实际感知到的浓度。因此,微区室化可以提供一种控制代谢途径的机制。与上述发展独立且并行的是,区室化的概念源于解释重要生理现象的必要性,特别是在心脏研究和心脏电生理学中。这些现象证明了本综述中描述的生物物理和生化机制的生理重要性。