CAMSAP2 organizes a γ-tubulin-independent microtubule nucleation centre

Microtubules are dynamic polymers consisting of αβ-tubulin heterodimers. The initial polymerization process, called microtubule nucleation, occurs spontaneously via αβ-tubulin. Since a large energy barrier prevents microtubule nucleation in cells, the γ-tubulin ring complex is recruited to the centrosome to overcome the nucleation barrier. However, detachment of a considerable number of microtubules from the centrosome is known to contribute to fundamental processes in cells. Here, we present evidence that minus-end-binding calmodulin-regulated spectrin-associated protein 2 (CAMSAP2) serves as a strong nucleator for microtubule formation from soluble αβ-tubulin independent of γ-tubulin. CAMSAP2 significantly reduces the nucleation barrier close to the critical concentration for microtubule polymerization by stabilizing the longitudinal contacts among αβ-tubulins. CAMSAP2 clusters together with αβ-tubulin to generate nucleation intermediates, from which numerous microtubules radiate, forming aster-like structures. Our findings suggest that CAMSAP2 supports microtubule growth by organizing a nucleation centre as well as by stabilizing microtubule nucleation intermediates.

Interaction of humic acids with soil minerals: adsorption and surface aggregation induced by Ca2+

Environmental context Humic acids, important components of natural organic matter in soils, sediments and aquatic media, can interact with the surface of minerals affecting key environmental processes. In the presence of calcium, humic acids can also interact among themselves leading to molecular aggregates. We demonstrate that a solid mineral surface facilitates the formation of humic acid aggregates, and thus surface aggregation occurs under conditions where normal aggregation in solution does not occur. Abstract Humic acids (HAs) interact with the surface of mineral particles leading to the formation of clay–humic complexes that affect the transport of nutrients and contaminants in the environment, soil structure, soil erosion and carbon sequestration by soils. The interaction is influenced by the presence of multivalent ions, such as Ca2+, which enhances the uptake of HAs by the particles. This article reports the effects of Ca2+ on the interaction between a HA and a soil clay fraction, both obtained from the same soil sample. The study was performed by using zeta potential measurements, HA adsorption isotherms, Ca2+ adsorption isotherms and microscopy. The results show that at low HA concentrations and low Ca2+ concentrations HA adsorption takes place, but that at high concentrations surface aggregation and precipitation also takes place, a process that is seldom reported or analysed in the literature. HA adsorption isotherms only give the overall HA uptake by the solid but they do not allow differentiation of HA adsorption from surface aggregation. However, HA adsorption v. Ca2+ concentration plots and Ca2+ adsorption isotherms at different HA concentrations can distinguish these two processes quite clearly. In addition, surface aggregation could be undoubtedly observed with optical microscopy. Surface aggregation starts to take place at a 0.7mM Ca2+ concentration, which is lower than the Ca2+ concentration needed to start HA aggregation in solution. This indicates that the surface of soil minerals acts as a nucleation centre for HA aggregation.

Dislocation Nucleation in Heteroepitaxial Semiconducting Films

The nucleation of dislocation in semiconductors is still a matter of debate and especially in heteroepitaxial films. To understand this nucleation process the classical models of dislocation nucleation are presented and discussed. Two main points are then developed: emission of dislocations from surface steps and the role of point defects agglomeration on dislocation nucleation. Recent atomic simulation of half loops emission from surface steps and experimental evidences of anisotropic relaxation of GaInAs films deposited on vicinal (111) GaAs substrates strongly support surface steps as preferential sites for nucleation. In low temperature buffer layer structures (SiGe/Si) an original dislocation structure is observed which corresponds to the dislocation emission in different glide systems by a unique nucleation centre.

SV40 AssemblyIn VivoandIn Vitro

The Simian virus 40 (SV40) capsid is aT = 7dicosahedral lattice ∼45 nm in diameter surrounding the ∼5 kb circular minichromosome. The outer shell is composed of 360 monomers of the major capsid protein VP1, tightly bound in 72 pentamers. VP1 is a jellyroll β-barrel, with extending N- and C-terminal arms. The N-terminal arms bind DNA and face the interior of the capsid. The flexible C-arms tie together the 72 pentamers in three distinct kinds of interactions, thus facilitating the formation of aT = 7 icosahedron from identical pentameric building blocks. Assemblyin vivowas shown to occur by addition of capsomers around the DNA. We apply a combination of biochemical and genetic approaches to study SV40 assembly. Ourin vivoandin vitrostudies suggest the following model: one or two capsomers bind at a high affinity toses, the viral DNA encapsidation signal, forming the nucleation centre for assembly. Next, multiple capsomers attach concomitantly, at lower affinity, around the minichromosome. This increases their local concentration facilitating rapid, cooperative assembly reaction. Formation of the icosahedron proceeds either by gradual addition of single pentamers to the growing shell or by concerted assembly of pentamer clusters.

Copper(II)-induced self-oligomerization of α-synuclein

α-Synuclein is a component of the abnormal protein depositions in senile plaques and Lewy bodies of Alzheimer's disease (AD) and Parkinson's disease respectively. The protein was suggested to provide a possible nucleation centre for plaque formation in AD via selective interaction with amyloid β/A4 protein (Aβ). We have shown previously that α-synuclein has experienced self-oligomerization when Aβ25-35 was present in an orientation-specific manner in the sequence. Here we examine this biochemically specific self-oligomerization with the use of various metals. Strikingly, copper(II) was the most effective metal ion affecting α-synuclein to form self-oligomers in the presence of coupling reagents such as dicyclohexylcarbodi-imide or N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline. The size distribution of the oligomers indicated that monomeric α-synuclein was oligomerized sequentially. The copper-induced oligomerization was shown to be suppressed as the acidic C-terminus of α-synuclein was truncated by treatment with endoproteinase Asp-N. In contrast, the Aβ25-35-induced oligomerizations of the intact and truncated forms of α-synuclein were not affected. This clearly indicated that the copper-induced oligomerization was dependent on the acidic C-terminal region and that its underlying biochemical mechanism was distinct from that of the Aβ25-35-induced oligomerization. Although the physiological or pathological relevance of the oligomerization remains currently elusive, the common outcome of α-synuclein on treatment with copper or Aβ25-35 might be useful in understanding neurodegenerative disorders in molecular terms. In addition, abnormal copper homoeostasis could be considered as one of the risk factors for the development of disorders such as AD or Parkinson's disease.

The sorption of zinc species by clay minerals

The effect of changes in pH and the presence of ligands on the uptake of zinc ions by three types of clay mineral (kaolinite, illite and montmorillonite) has been investigated. In alkaline media the clay suspension acts as a nucleation centre for polymeric hydroxy species, and the major role of many ligands is to mask the precipitation process. Uncharged and negatively charged species are not sorbed to any measurable extent. In acidic media the adsorption capacity of the clays for zinc increases with pH and possible mechanisms are considered. For kaolinite and illite the controlling process appears to be the attachment of hydroxy species to particular sites on the particle edges; with montmorillonite ion exchange at negative lattice sites appears predominant. Of particular interest is the apparent affinity between montmorillonite and species containing nitrogen functional groups.

The sorption of copper species by clays. I. Kaolinite

The effect of changes in pH and the presence of ligands on the uptake of copper ions by kaolin has been investigated. In alkaline media the clay suspension acts as a nucleation centre for hydroxy-bridged copper species, and the major role of many ligands is to 'mask' this precipitation reaction. Uncharged and negatively charged complex species are not sorbed to any measurable extent. In acidic media, the adsorption capacity of the clay for cationic species increases with pH. Probable mechanisms are discussed and the feasibility of predicting the effects of ligands on sorption considered.