Prediction of location of active sites in biologically active peptides

In a previous paper we demonstrated that the short-range compact regions in atrial natriuretic factor (-hANF) predicted by the average distance map (ADM) correspond to its active sites [Kikuchi,J. Protein Chem.11, 579–581 (1992)]. In the present paper we apply the same method to other bioactive peptides and peptidic enzyme inhibitors. We again observe that active sites in each peptide are contained in short-range compact regions predicted by the ADM for the peptide. This demonstrates that the ADM method predicts the possible location of active sites in biologically active peptides in general. The possibility of practical application of the present method to rational drug design is also discussed.     

Antibacterially active substances

Synthesis of two hydroxy-derivatives of nalidixic acid as a result of microbial transformation was demonstrated in certain species of the genusAspergillus. Aspergillus alliaceus produced 7-hydroxy-nalidixic acid andAspergillus niger 6-hydroxy-nalidixic acid. It was demonstrated that the antibacterial activity of both hydroxy-derivatives (tested inEscherichia coli) was lower than that of the initial nalidixis acid.     

生物活性肽

生物活性肽是对生物机体的生命活动有益或具有生理作用的肽类化合物,简述了生物活性肽的分类、特点、作用机理以及开发应用。     

Antimicrobially active substances

The antimycotic activity of 41 quaternary ammonium compounds was studied. The spectrum of their effect was evaluated in eight microbial strains. Homologues, isomers or close analogues of Septonex, i.e. α-carbetoxypentadecyl-trimethylammonium bromide (XII) were found to be the most active. The presence of an alicyclic or an aromatic ring or of a pyridine residue inhibited antimycotic activity, while substitution of hydrogen by a halogen potentiated it.     

Secondary active transport

Secondary active transport is defined as the transport of a solute in the direction of its increasing electrochemical potential coupled to the facilitated diffusion of a second solute (usually an ion) in the direction of its decreasing electrochemical potential. The coupling agents are membrane proteins (carriers), each of which catalyzes simultaneously the facilitated diffusion of the driving ion and the active transport of a given solute. The review starts with some considerations on the energetics followed by a presentation of the kinetics of secondary active transport. Examples of information which may be gained by such studies are discussed. In the second part, some examples of secondary transport are given; we also describe the characteristics of the corresponding carriers. The various transport systems presented are: the D-glucose/Na+ symport in brush-border membranes, the lactose/H+ symport in E. coli, the Na+/H+ antiport, the different transport systems in the inner mitochondrial membrane.     

Bacterial intracellularly active toxins: Membrane localisation of the active domain

Numerous bacterial toxins exert their activity by inactivating or modulating a specific intracellular host target. For this purpose, these toxins have developed efficient strategies to overcome the different host cell defences including specific binding to cell surface, internalisation, passage through the endosome or plasma membrane, exploiting intracellular trafficking and addressing to intracellular targets. Several intracellularly active toxins deliver an active domain into the cytosol that interacts with a target localised to the inner face of the plasma membrane. Thus, the large clostridial glucosylating toxins (LCGTs) target Rho/Ras‐GTPases, certain virulence factors of Gram negative bacteria, Rho‐GTPases, while Pasteurella multocida toxin (PMT) targets trimeric G‐proteins. Others such as botulinum neurotoxins and tetanus neurotoxin have their substrate on synaptic vesicle membrane. LCGTs, PMT, and certain virulence factors from Vibrio sp. show a particular structure constituted of a four‐helix bundle membrane (4HBM) protruding from the catalytic site that specifically binds to the membrane phospholipids and then trap the catalytic domain at the proximity of the membrane anchored substrate. Structural and functional analysis indicate that the 4HBM tip of the Clostridium sordellii lethal toxin (TcsL) from the LCGT family contain two loops forming a cavity that mediates the binding to phospholipids and more specifically to phosphatidylserine.     

Interference of active site specific reagents in plasminogen-streptokinase active site formation

We have recently observed slow, non-Michaelis-Menten kinetics of activation of native cat plasminogen by catalytic concentrations of streptokinase. In order to understand the reasons for this phenomenon, we undertook to study the formation of the plasminogen-streptokinase activator complex under the same plasminogen activation conditions. The results obtained in this study show that the potential active site in both cat and human plasminogen is capable of binding strongly the specific substrates (S) p-nitrophenyl p-guanidinobenzoate (NPGB) and H-D-valyl-L-leucyl-L-lysyl-p-nitroanilide, through the active site is incapable of hydrolyzing these substrates. Binding studies support these and the following conclusions. Streptokinase binds to this zymogen-substrate complex to create the ternary plasminogen-S-streptokinase complex, which then slowly converts to an acylated plasminogen-streptokinase form. This acylation reaction is 550 times slower than acylation of the preformed plasminogen-streptokinase complex by NPGB. The same reaction also occurs with human plasminogen, though the acylation reaction is 10 times faster than when the cat zymogen is used. NPGB binds specifically to plasminogen but not to streptokinase. These studies proved that inhibition of cat plasminogen activation by streptokinase occurs at the level of activator complex formation. We conclude from our studies that streptokinase binding to both cat and human plasminogen occurs at the potential active site of the zymogen. Consequently, it is probable that plasminogen activation in vivo is inhibited by binding of active site specific inhibitors to plasminogen.     

Rhamnogalacturonan-II--a biologically active fragment

Rhamnogalacturonan-II inhibited the uptake of [¹⁴C]leucine and,consequently, the incorporation of [¹⁴C]leucine into acid-precipitableproteins by suspension-cultured tomato cells. Fractionationof rhamnogalacturonan-II showed that the lower molecular componentswere the most effective. KDO and apiose, both constituents ofrhamnogalacturonan-II, also inhibited [¹⁴C]leucine incorporationweakly, suggesting that these sugar residues may be an integralrequirement for the biological activity of rhamnogalacturonan-II.The incorporation of [¹⁴C]glutamate and [¹⁴C]histidine, andto a lesser extent [¹⁴C]proline and [¹⁴C]arginine, was alsoinhibited by rhamnogalacturonan-II; the incorporation of [¹⁴C]tyrosineand [¹⁴C]phenylalanine was little affected. This suggests thatrhamnogalacturonan-II exerts its effect by acting on certainmembrane transport systems. Key words: Rhamnogalacturonan-II, inhibition, protein synthesis, amino acid incorporation     

Morphogenesis of active shells

We consider the active shell as a single-cell or epithelial sheet surface that, sharing basic properties of stretched elastic shells, is capable of active planar movement owing to recruiting of the new surface elements. As model examples of their morphogenesis, we consider the growth and differentiation of single-cell hairs (trichomes) in plants of the genus Draba, and the epiboly and formation of the dorsoventral polarity in loach. The essential feature of the active shell behavior at both cellular and supracellular levels is regular deviating from the spatially homogeneous form, which is a primary cause of originating of the active mechanical stresses inside the shell in addition to its passive stretching by the intrinsic forces. Analyzing the quantitative morphological data, we derive the equations in which the temporal self-oscillations and spatial differentiation are distinguishable only at the parametric level depending on the proportion of active to passive stresses. In contrast to the ordinary activator-inhibitor systems, the self-oscillation dynamics is principally non-local and, consequently, one-parametric, the shell surface curvature being an analog of the inhibitor, while its spatial variance being an analog of the activator of shaping. Analyzing variability and evolution of the hair cell branching, we argue that the linear ontogeny (succession of the developmental stages) is a secondary evolutionary phenomenon originating from cyclic self-organizing algorithms of the active shell shaping.     

The presynaptic active zone

Neurotransmitters are released by synaptic vesicle exocytosis at the active zone of a presynaptic nerve terminal. In this review, I discuss the molecular composition and function of the active zone. Active zones are composed of an evolutionarily conserved protein complex containing as core constituents RIM, Munc13, RIM-BP, α-liprin, and ELKS proteins. This complex docks and primes synaptic vesicles for exocytosis, recruits Ca(2+) channels to the site of exocytosis, and positions the active zone exactly opposite to postsynaptic specializations via transsynaptic cell-adhesion molecules. Moreover, this complex mediates short- and long-term plasticity in response to bursts of action potentials, thus critically contributing to the computational power of a synapse.