Conversion of albumin into a transmitter of the ultra long-range interaction of human erythrocytes

Previous experiments in our laboratory and elsewhere have shown that extended macromolecules of high molecular weight are needed in the suspending medium to transmit the quantum-mechanical coherent interaction of human erythrocytes. Albumin, a globular protein, does not transmit the interaction at physiological concentrations. Albumin can be converted into a transmitter by preheating a solution of albumin to 62 degrees C for 20 min. Such treatment unravels the tertiary structure of albumin and allows it to polymerize.     

Conversion of albumin into a transmitter of the ultra long-range interaction of human erythrocytes

Previous experiments in our laboratory and elsewhere have shown that extended macromolecules of high molecular weight are needed in the suspending medium to transmit the quantum-mechanical coherent interaction of human erythrocytes. Albumin, a globular protein, does not transmit the interaction at physiological concentrations. Albumin can be converted into a transmitter by preheating a solution of albumin to 62°C for 20 min. Such treatment unravels the tertiary structure of albumin and allows it to polymerize.     

荧光能量转移技术在生物学研究中的应用

荧光能量转移(FRET)是指两个携带不同荧光基团的大分子在相互间距离足够近时(10～100A)所发生的能量非放射性地由一个荧光基团向另一个荧光基团转移的现象。结合绿色荧光蛋白的发现,FRET技术可用于检测生物大分子中不同亚基的位置和生物大分子间的相互作用。近年来,FRET技术在生物学研究中的突破性进展是在活体细胞中实时监测生物大分子之间的相互作用。本文就绿色荧光蛋白的发现,FRET技术的原理、研究进展和应用前景作简要综述。     

Tyrosine phosphorylation and the mechanism of signal transduction by the B-lymphocyte antigen receptor.

Lymphocytes provide a powerful defense against infectious agents with their exquisite ability to distinguish between macromolecules of the host and macromolecules of foreign invaders. This ability derives from the antigen receptors, which are created from precursor minigenes by a series of genetic-recombination reactions [1, 2] and from cellular mechanisms that inactivate lymphocytes expressing self-reactive antigen receptors [3, 4]. Central to the problem of distinguishing self from non-self is the means by which these antigen receptors recognize antigen and transmit the information of that recognition to the interior of the cell. This information ultimately leads to lymphocyte activation or inactivation, depending upon the context. In this review, I shall summarize recent advances in understanding the structural elements of the antigen receptor complex of B lymphocytes and in understanding the signal-transduction events initiated by this receptor.     

Mobile Macromolecules in Plant Development

Plant cells transmit developmental signals and distribute nutrients via dynamic intercellular channels termed plasmodesmata (PD). Multidisciplinary inquiries have provided evidence that plasmodesmatal regulation is critical to fundamental plant functions, such as development, host–pathogen interactions, and systemic RNA silencing. This review focuses on macromolecules that transport cell-to-cell through PD and describes their implications on plant development.     

Kerr effect of charged dipolar macromolecules without condensed counterions in conducting solution.

The theoretical treatment of the Kerr constant of rigid, dipolar, conducting ellipsoidal macromolecules of O'Konski and Krause (1970. J. Phys. Chem. 74:3243) has been extended to very low ionic strength solutions for charged macromolecules. The O'Konski and Krause theoretical treatment postulated a surface conductivity directly on the surface of each macromolecule. For charged macromolecules, this surface conductivity was generally assumed to be caused by movement of condensed counterions on the macromolecules. In the present work, it has been assumed that, at very low ionic strength, the average counterion is at the Debye characteristic distance from the surface of each charged macromolecule and contributes to surface conductivity at that distance, with no additional surface conductivity on the true surface of the macromolecule. Essentially, these considerations change the calculated interaction energy of the macromolecule with an externally applied electric field via a change in both the internal field components and in the reaction field of the macromolecular dipole. The new interaction energy is used to calculate the orientation distribution function of the macromolecules in solution and this distribution function can, in principle, be used to calculate the steady state electric linear or circular dichroism, electric light scattering, anisotropy of conductivity, etc., using the appropriate theoretical treatment for each of these quantities.     

1H magnetization transfer in hydrated gluten and flour: effects of wheat aging

The interaction of water with flour or gluten in hydrated samples was investigated by proton magnetization transfer measurements. Flour and gluten from both durum and bread wheat seeds, either unaged or artificially aged over different periods of time, were investigated. Measurements were performed at several radio frequency power levels and frequency offsets, and the data were quantitatively modeled by two interacting pools, a liquid (water) and a solid (macromolecules) one. A super-Lorentzian line shape well described the magnetization of the solid pool. Magnetization transfer was found to be more efficient for flour with respect to gluten samples, in agreement with their hydrophilic/hydrophobic behavior. The aging treatment of seeds resulted in a minor degree of interaction between macromolecules and water.     

Boundary element solution of macromolecular electrostatics: interaction energy between two proteins.

The boundary element technique is implemented to solve for the electrostatic potential of macromolecules in an ionic solution. This technique entails solving surface integral equations that are equivalent to the Poisson and the Poisson-Boltzmann equations governing the electrostatic potential inside the macromolecules and and in the solvent. A simple but robust method is described for discretizing the macromolecular surfaces in order to approximate the integral equations by linear algebraic equations. Particular attention is paid to the interaction energy between two macromolecules, and an iterative procedure is devised to make the calculation more efficient. This iterative procedure is illustrated in the electron transfer system of cytochrome c and cytochrome c peroxidase.     

Using Lamm-Equation modeling of sedimentation velocity data to determine the kinetic and thermodynamic properties of macromolecular interactions

The interaction of macromolecules with themselves and with other macromolecules is fundamental to the functioning of living systems. Recent advances in the analysis of sedimentation velocity (SV) data obtained by analytical ultracentrifugation allow the experimenter to determine important features of such interactions, including the equilibrium association constant and information about the kinetic off-rate of the interaction. The determination of these parameters is made possible by the ability of modern software to fit numerical solutions of the Lamm Equation with kinetic considerations directly to SV data. Herein, the SV analytical advances implemented in the software package SEDPHAT are summarized. Detailed analyses of SV data using these strategies are presented. Finally, a few highlights of recent literature reports that feature this type of SV data analysis are surveyed.     

A method of reversible biomolecular immobilization for the surface plasmon resonance quantitative analysis of interacting biological macromolecules

This article presents a new procedure for the immobilization of macromolecules on gold surfaces, with the purpose of studying macromolecular interactions by simple optical configurations rendering surface plasmon resonance. Gold surfaces were covered by a three-layer structure composed of poly-L-lysine irreversibly bound to gold, followed by a second layer of heparin and a third layer of polylysine. The three-layer structure of polylysine-heparin-polylysine remains irreversibly bound to gold, it prevents biomolecules from coming into direct contact with the metal surface, and it allows the irreversible binding of different proteins and polynucleotides. After binding of a macromolecule to the three-layer structure, the interaction with a second macromolecule can be studied, and then the complex formed by the two interacting macromolecules, together with the second heparin layer and the third polylysine layer, can be broken down just by treatment with an alkaline solution having a pH value above the pK value of the amino groups of polylysine. The first polylysine layer remains irreversibly bound to gold, ready to form a new three-layer structure and, therefore, to support a new macromolecular interaction on the same regenerated surface. Polynucleotide interactions, the proteolytic action of chymotrypsin, and the interaction between the component subunits of a heterotetrameric enzyme are described as examples of macromolecular interactions studied by using this system. The method may be especially suitable for developing of low-cost systems aimed to look for surface resonance signals, and it offers the advantage of allowing calculation of parameters related to the size and stoichiometry of the interacting macromolecules, in addition to the kinetic and equilibrium properties of the interaction.     

Histochemical properties of cartilage proteoglycans

Proteoglycan interaction with alcian blue at different concentrations of magnesium chloride was studied both in vitro and in histological sections of paraffin-embedded tissues. Our experiments indicate that a) proteoglycans with different contents of chondroitin sulfate and keratan sulfate, prepared under nondegradative conditions, are not distinguishable on the basis of the critical electrolyte concentrations at which staining is abolished; b) the state of aggregation of proteoglycans only very slightly affects the alcian blue affinity of the macromolecules at different concentrations of magnesium chloride; c) the interaction of proteoglycans with other components of the connective tissue matrix is an important factor in determining the strength of binding of alcian blue to the polyanionic macromolecules in histological sections. These factors should be considered in interpreting histochemical data obtained by staining tissue sections with alcian blue at different concentrations of magnesium chloride. Proteoglycans, like glycosaminoglycans, are only weakly periodic acid-Schiff-positive.     

Thermal resistance in fungi: the role of heat shock proteins and trehalose

The parallel synthesis of heat shock proteins and trehalose in response to heat shock did not allow the role of these compounds in the acquisition of thermotolerance by fungal cells to be established for a long time. This review analyses experimental data obtained with the use of mutant fungal strains and shows differences in the thermoprotective functions of trehalose and heat shock proteins in relation to cell membranes and macromolecules. The main emphasis has been placed on data demonstrating the thermoprotective role of trehalose in fungi, the present-day understanding of its biological functions, and mechanisms of trehalose interaction with subcellular structures and cell macromolecules.     

Isolation of cloned variants of a rat hepatoma cell line with altered attachment to collagen, but normal attachment to fibronectin

Cloned variants of a rat hepatoma cell line have been isolated which exhibit normal attachment and spreading behavior on fibronectin substrata, but which are defective in their ability to attach to native collagen films. These clones should be useful for identifying specific macromolecules involved in the cell-to-collagen interaction.     

Mapping of isozymic differences in enolase

The existence of the isozymes of non-regulatory enzymes often has been linked to their interaction with other macromolecules. Enolase, a non-regulatory enzyme, has three isozymes for which sequences have been determined in two or more vertebrate species. The positions in the enolase sequences that differ between the isozymes were mapped in the 3-D structure of the enzyme. The positions in a given isozymic form which were not conserved in different species were considered to be resulting from the neutral drift of sequences and rejected. Also, the residues with no accessible surface were rejected. Three areas with relatively high densities of isozymic substitutions were found. We consider them as the likely sites of contact with other macromolecules.     

Proteins and vesicular transport in capillary endothelium

Plasma proteins interact with vascular endothelium in such a way as to render it less permeable to other macromolecules. Evidence from a variety of sources indicates that this may result from interaction of the circulating macromolecules with the negatively charged glycoprotein layer on the surface of endothelial cells, and that this layer may be responsible for some of the known molecular sieving properties attributed to the endothelium. Experiments with the fluorocarbon exchange-transfused rat are described, which suggest that there may be mechanisms other than vesicular translocation that facilitate the passage of macromolecules across endothelium. Such mechanisms include, among others, the formation of transient transendothelial channels that appear to be less sensitive than pinocytotic vesicles to the concentration of ambient protein. Recent evidence suggests that, in addition to molecular size and charge, glycosylation of protein molecules and cell membranes themselves may facilitate vesicular uptake.     

The interaction of plasminogen activator with a reconstituted basement membrane matrix and extracellular macromolecules produced by cultured epithelial cells

Laminin and fibronectin are glycoproteins that influence cell behavior and mediate cell/substratum adhesion. We have examined the interaction of these macromolecules with the serine protease plasminogen activator (PA) in two types of extracellular matrices; one produced by the murine Engelbreth-Holm-Swarm (EHS) tumor (Matrigel), and another by normal kidney epithelial cells in culture. Matrigel was found to contain significant quantities of tissue-type PA (tPA). Two of the major components of Matrigel, laminin and type IV collagen, were also examined. Tissue-type PA was associated with purified preparations of laminin; however, it was not found associated with type IV collagen. Normal kidney epithelial cells in culture secrete large amounts of urokinase (UK) and deposit a subepithelial matrix containing both laminin and fibronectin. These matrix macromolecules were isolated from the deposited matrix by immunoprecipitation, examined by zymography, and found to contain UK. The potential role of this interaction in the mechanisms of cell migration and matrix remodeling is discussed.     

蛋白质分子识别新认识

近几年,国际上肽库技术得到了迅速的发展,在抗原决定簇的定位及分子识别方面积累了大量的数据．就近3年来蛋白质分子识别方面的观念和认识上的深化进行了综述：a．蛋白质间的相互作用是少数几个关键残基的弱相互作用提供了大部分结合能;b．这种相互作用可以用小肽来模拟．因此通过研究小肽片段的特异性相互作用,可以揭示蛋白质间相互作用的本质,为小肽分子药物和疫苗的设计,乃至复杂超分子体系的研究提供了理论基础和实验依据．     

Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm

Bidirectional transport of macromolecules between the nucleus and the cytoplasm occurs through the nuclear pore complexes (NPCs) by a signal-mediated mechanism that is directed by targeting signals (NLSs) residing on the transported molecules or "cargoes." Nuclear transport starts after interaction of the targeting signal with soluble cellular receptors. After the formation of the cargo-receptor complex in the cytosol, this complex crosses the NPC. Herein, we use gold particles of various sizes coated with cargo-receptor complexes to determine precisely how large macromolecules crossing the NPC by the signal-mediated transport mechanism could be. We found that cargo-receptor-gold complexes with diameter close to 39 nm could be translocated by the NPC. This implies that macromolecules much larger than the assumed functional NPC diameter of 26 nm can be transported into the karyoplasm. The physiological relevance of this finding was supported by the observation that intact nucleocapsids of human hepatitis B virus with diameters of 32 and 36 nm are able to cross the nuclear pore without disassembly.     

The role of encapsulation by β-cyclodextrin in the interaction of raloxifene with macromolecular targets: a study by spectroscopy and molecular modeling

We report the binding of the drug raloxifene with Calf thymus DNA (ctDNA) and bovine serum albumin (BSA) in the presence and absence of β-cyclodextrin (β-CD) and explain the influence of β-cyclodextrin on the binding of the drug to macromolecules. UV-Vis absorption, fluorescence, proton nuclear magnetic resonance and two-dimensional rotating-frame nuclear overhauser effect spectroscopic techniques are used to study the stoichiometry and the binding strength of the complexes. Molecular modeling is used in combination with other techniques to propose the structure of the inclusion complex and the interaction with ctDNA. The Stern–Volmer quenching constants of the interaction of raloxifene with ctDNA in aqueous and in β-CD solution are compared. The competition for binding of ctDNA with raloxifene and Methylene Blue is studied. The apparent binding constant and the number of binding sites for the binding of raloxifene with BSA in aqueous solution are significantly different from those in the presence of β-CD. The influence of β-CD on the binding of the small molecules with biological macromolecules is discussed. We infer that the binding strengths between raloxifene and macromolecules, viz., ctDNA and BSA are influenced by the β-CD encapsulation. These results may suggest new ways to tune the drug binding to biomacromolecules by encapsulating specific moieties of drugs.     

Detection of living cervical cancer cells by transient terahertz spectroscopy

The photonic energy of terahertz wave is in the same order of magnitude as the rotational and vibrational energy levels of organic and biological macromolecules, so it has unique advantages in detecting cells and biological macromolecules. However, in the life environment, the dynamic time scale of cell-environment interaction and structural conformation change of biological macromolecules are within picosecond to millisecond, and water has strong absorption to terahertz wave, which has become the bottleneck problem for the detection of cells and biological macromolecules by terahertz technology. In this article, we developed a set of terahertz single measurement system based on the tilt wave front of grating pulse technique. The system was employed for the terahertz detection of trace living cervical cancer cells. We achieved transient detection of the terahertz pulse time-domain waveform of the living HeLa cells. The characteristic absorption peaks were identified by Lambert-Beer law, respectively, at 0.49, 0.71, 1.04, 1.07, 1.26 and 1.37 THz. The absorbance is proportional to the cell concentration.