Self-assembly microtubule systems

Self-assembling systems made of proteins provide many biological systems with essential structural elements, including viral envelopes, bacterial S-layers, microtubules, collagens, and keratins (see Ref. 6 for a review). Detailed studies on in vitro self-assembly optimization were carried out for several systems as a part of an ongoing effort to elucidate the in vivo mechanism. Data reported from such studies may provide an optional basis for the design and in vitro fabrication of nanostructures made of natural proteins (Fig. 1). 

The polymer self-assembly theory of Oosawa and Kasai (1962) provides valuable insights into the nature of the nucleation process. The polymerization nucleus is considered to form by the accretion of protomers, but the process is highly cooperative and unfavorable. Indeed, this is strongly suggested by the observation that thousands of actin or tubulin protomers are found in F-actin and microtubule structures if nucleation of self-assembly were readily accomplished and highly favorable, the consequence would be that many more fibers of shorter polymer length would be observed. The Oosawa kinetic theory for nucleation permits one to obtain information about the size of the polymerization nucleus if two basic assumptions can be satisfied in the experimental system. First, the rate of nuclei formation is assumed to be proportional to the loth power of the protomer concentration with io representing the number of protomers required to create the nucleus. Second, the treat-... 

As noted earlier, microtubule elongation has been characterized largely with respect to the involvement of guanine nucleotides and the modes of drug inhibition of microtubule formation. There have also been a number of important studies on the influence of microtubule-associated proteins and solution variables on the kinetics and thermodynamics of microtubule self-assembly. Of these, the characterization of the so-called mitotic spindle poisons has been particularly complex because of the variety of agents and the diversity of systems studied. For this reason, we shall concentrate on the other factors affecting the elongation process. 

The threshold concentration of monomer that must be exceeded for any observable polymer formation in a self-assembling system. In the context of Oosawa s condensation-equilibrium model for protein polymerization, the cooperativity of nucleation and the intrinsic thermodynamic instability of nuclei contribute to the sudden onset of polymer formation as the monomer concentration reaches and exceeds the critical concentration. Condensation-equilibrium processes that exhibit critical concentration behavior in vitro include F-actin formation from G-actin, microtubule self-assembly from tubulin, and fibril formation from amyloid P protein. Critical concentration behavior will also occur in indefinite isodesmic polymerization reactions that involve a stable template. One example is the elongation of microtubules from centrosomes, basal bodies, or axonemes. 

There are several self-assembling macromolecules that are of interest to us in this text. They include (1) collagen, the primary structural material found in the extracellular matrix (2) actin, a component of the cell cytoskeleton that is involved in cell locomotion and in formation of the thin filaments of muscle (3) microtubules, which are involved in cell mitosis, movement, and organelle movement and finally (4) fibrinogen, which forms fibrin networks that minimize bleeding from cut vessels. Self-assembly is important in these systems because the function of these macromolecules can be modified via processes that increase the molecular axial ratio and hence decrease the solubility. 

The self-assembly of tubulin to form microtubules was described initially in a classic polymerization model of nucleated helical polymerization by Maruyama and Oosawa (4). Assembly involves two phases a nucleation phase followed by an elongation phase. With purified systems in vitro, nucleation can... 

Here, as elsewhere in cell biology, rich interconnections between problems have long been obvious or may now be inferred. In this review, some attention is given to the relations between mitotic phenomena and these other areas of study reversible self-assembly of protein aggregates, other motile systems and the burgeoning information on microtubules, interactions between DNA and proteins, and the recently discovered, very different chromosome motion and distribution mechanisms in lower organisms. 

Nanostmcture science and supramolecular chemistry are fast evolving fields that are concerned with manipulation of materials that have important structural features of nanometer size (1 run to 1 pm) [10, 11]. Nature has been exploiting no covalent interactions for the construction of various cell components. For instance, microtubules, ribosomes, mitochondria, and chromosomes use mostly hydrogen bonding in conjunction with covalently formed peptide bonds to form specific stmctures. The self-assembled systems and self-organized stmctures mediated by transition metals are considered in coimection with increasing research interest in chemical transformations with use of these systems [12]. 

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