Membrane fusion

Summary The factors involved in the regulation of biological membrane fusion and models proposed for the molecular mechanism of biomembrane fusion are reviewed. The results obtained in model systems are critically discussed in the light of the known properties of biomembranes and characteristics of biomembrane fusion. Biological membrane fusion is a local-point event; extremely fast, non-leaky, and under strict control. Fusion follows on a local and most probably protein-modulated destabilization, and a transition of the interacting membranes from a bilayer to a non-bilayer lipid structure. The potential role of type II non-bilayer preferring lipids and of proteins in the local destabilization of the membranes is evaluated. Proteins are not only responsible for the mutual recognition of the fusion partners, but are most likely also to be involved in the initiation of biomembrane fusion, by locally producing or activating fusogens, or by acting as fusogens.     

Membrane fusion: the process and its energy suppliers

Membrane fusion constitutes a pivotal process in eukaryotic cell physiology. Both specialized proteins and membrane lipids play key roles in fusion. Here, our current understanding of the mechanism of membrane fusion is reviewed. The focus is on the relatively simple and well-understood proteinaceous fusion machinery of enveloped viruses and the physical properties of lipids that appear to be of great relevance for fusion progression. Recent observations suggest that viral fusion proteins use packed conformational energy and bilayer-destabilizing domains to (i) bring participating membranes into intimate contact, (ii) merge proximal lipid monolayers through highly curved stalk/hemifusion intermediates, and (iii) generate a lipid-containing fusion pore, thereby terminating the fusion process. Received 4 January 2002; received after revision 3 April 2002; accepted 5 April 2002     

The Membrane Protein Data Bank

The Membrane Protein Data Bank (MPDB) is an online, searchable, relational database of structural and functional information on integral, anchored and peripheral membrane proteins and peptides. Data originates from the Protein Data Bank and other databases, and from the literature. Structures are based on X-ray and electron diffraction, nuclear magnetic resonance and cryoelectron microscopy. The MPDB is searchable online by protein characteristic, structure determination method, crystallization technique, detergent, temperature, pH, author, etc. Record entries are hyperlinked to the PDB and Pfam for viewing sequence, three-dimensional structure and domain architecture, and for downloading coordinates. Links to PubMed are also provided. The MPDB is updated weekly in parallel with the Protein Data Bank. Statistical analysis of MPDB records can be performed and viewed online. A summary of the statistics as applied to entries in the MPDB is presented. The data suggest conditions appropriate for crystallization trials with novel membrane proteins. Received 3 August 2005; received after revision 18 September 2005; accepted 26 September 2005 This paper and the Membrane Protein Data Bank celebrate the 20^th anniversary of the landmark paper in Nature (1985, 318: 618–624) describing the first ‘high-resolution’ three-dimensional structure of a membrane protein, the photosynthetic reaction center from Rhodopseudomonas (Blastochloris) viridis.     

Membrane biology and signal transduction

Posters

Membrane biology and signal transduction     

High-frequency fusion of fungal protoplasts

Zusammenfassung Es gelang, die intraspezifische Protoplastfusion vonAspergillus flavus, A. nidulans, A. niger, Penicillium frequentans undP. ramigena mit grosser H?ufigkeit zu verwirklichen. Die Fusionsh?ufigkeit wurde durch Komplementation von Mangelmutanten festgestellt.

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The authors thank Dr.M. Czakó, Dr.M. Pesti and Mr.L. Manczinger for producing and characterizing some of the required mutants, andéva Sziráki andMagdolna Romvári for skilful technical assistance.     

Membrane traffic in the secretory pathway

Vesicular transport is the basic communication mechanism between different compartments in a cell and with the environment. In this review I discuss the principles of vesicle generation and consumption with particular emphasis on the different types of coat proteins and the timing of the shedding of the coat proteins from transport containers. In recent years it has become clear that there are more coat complexes than the classical COPI, COPII and clathrin coats. These additional coats may generate vesicles that transport cargo in a temporally and/or spatially controlled manner. Work over the last years suggests that GTP hydrolysis occurs early during vesicle biogenesis, destabilizing the coat perhaps before fission of the vesicle from the donor membrane occurs. Recent findings imply, however, that tethers at the receiving compartment specifically detect the coat on vesicle. (Part of a Multi-author Review)     

Membrane traffic in the secretory pathway

Vesicular transport is the basic communication mechanism between different compartments in a cell and with the environment. In this review I discuss the principles of vesicle generation and consumption with particular emphasis on the different types of coat proteins and the timing of the shedding of the coat proteins from transport containers. In recent years it has become clear that there are more coat complexes than the classical COPI, COPII and clathrin coats. These additional coats may generate vesicles that transport cargo in a temporally and/or spatially controlled manner. Work over the last years suggests that GTP hydrolysis occurs early during vesicle biogenesis, destabilizing the coat perhaps before fission of the vesicle from the donor membrane occurs. Recent findings imply, however, that tethers at the receiving compartment specifically detect the coat on vesicle. (Part of a Multi-author Review)     

Membrane traffic in the secretory pathway

Secretion is a fundamental biological activity of all eukaryotic cells by which they release certain substances in the extracellular space. It is considered a specialized mode of membrane trafficking that is achieved by docking and fusion of secretory vesicles to the plasma membrane (i.e., exocytosis). Secretory vesicle traffic is thought to be regulated by a family of Rab small GTPases, which are regulators of membrane traffic that are common to all eukaryotic cells. Classically, mammalian Rab3 subfamily members were thought to be critical regulators of secretory vesicle exocytosis in neurons and endocrine cells, but recent genetic and proteomic studies indicate that Rab3 is not the sole Rab isoform that regulates secretory vesicle traffic. Rather, additional Rab isoforms, especially Rab27 subfamily members, are required for this process. In this article I review the current literature on the function of Rab isoforms and their effectors in regulated secretory vesicle traffic.     

Membrane currents in cardiac pacemaker tissue

The present work is a brief survey of the mechanism of the cardiac pacemaker in sinoatrial node cells. Information on the pacemaker mechanism in cardiac tissue has been greatly enhanced by the development of the single cell isolation technique and the patch clamp technique. These methods circumvent to a large extent the difficulties involved in voltage clamping multicellular preparations. The calcium current (ICa), delayed rectifier potassium current (IK), transient outward current (Ito;IA), and the hyperpolarization activated inward current (Ih or If) were found both in whole cell preparations and in single channel analysis. The physiological significance of these currents, together with the exchange current systems for the pacemaker depolarization are discussed.     

膜分离处理印染废水研究进展

印染废水具有水量大、色度高、成分复杂、环境污染严重等特点。膜分离技术处理印染废水具有选择性好、生产效率高和处理成本低等特点。基于对近年来的文献调研,综述了膜分离技术在印染废水处理中的研究进展情况,指出了膜分离法处理印染废水还存在的主要问题和未来发展方向,并对膜分离技术处理印染废水应用前景作了展望。     

Membrane traffic in the secretory pathway

Cholesterol, certain lipids, membrane-bound and soluble proteins, as well as viruses that are synthesized in the endoplasmic reticulum (ER), reach the plasma membrane (PM) via non-classical pathway(s) that remain poorly understood. Typical for this transport is (i) its insensitivity to brefeldin A (BFA), which dissociates selected coat complexes from membranes, resulting in the disassembly of the Golgi apparatus; (ii) its rapid kinetics as compared to the classical secretory pathway; and (iii) its role in the trafficking of lipid raft components. Based on results showing that the intermediate compartment (IC) at the ER-Golgi boundary constitutes a stable tubular network that maintains its dynamics in the presence of BFA, we propose that two bidirectional Golgi-bypass pathways to the PM exist, a direct route from early IC elements, and another, reminiscent of the yeast secretory pathway, from late IC elements via the endosomal system. These pathways have implications for the organization of the secretory processes in different cell types.     

Membrane traffic in the secretory pathway

Membrane trafficking is crucial in the homeostasis of the highly compartmentalized eukaryotic cells. This compartmentalization occurs both at the organelle level, with distinct organelles maintaining their identities while also intensely interchanging components, and at a sub-organelle level, with adjacent subdomains of the same organelle containing different sets of lipids and proteins.Acentral question in the field is thus how this compartmentalization is established and maintained despite the intense exchange of components and even physical continuities within the same organelle. The phosphorylated derivatives of phosphatidylinositol, known as the phosphoinositides, have emerged as key components in this context, both as regulators of membrane trafficking and as finely tuned spatial and temporal landmarks for organelle and sub-organelle domains. The central role of the phosphoinositides in cell homeostasis is highlighted by the severe consequences of the derangement of their metabolism caused by genetic deficiencies of the enzymes involved, and from the systematic hijacking of phosphoinositide metabolism that pathogens operate to promote their entry and/or survival in host cells. (Part of a Multi-author Review)     

Membrane traffic in the secretory pathway

The steady-state localisation of membrane proteins in the endocytic system is the result of many sorting events that occur at various points throughout the endosomal pathway. A protein that has been endocytosed from the plasma membrane or sorted at the trans-Golgi network (TGN) and transported to an endosome will ultimately be delivered to one of three destinations: the plasma membrane, the TGN or the lysosome. Where a membrane protein is trafficked to depends on the interactions between sorting motifs present in the membrane protein and the machinery that can decode these motifs. Much of the protein machinery that recognises sorting motifs is conserved from yeast toman, and in this review I will discuss this machinery and the motifs that govern endosomal protein sorting. (Part of a Multi-author Review)     

Membrane traffic in the secretory pathway

SNARE (SNAP receptor) proteins drive intracellular membrane fusion and contribute specificity to membrane trafficking. The formation of SNAREpins between membranes is spatially and temporally controlled by a network of sequentially acting accessory components. These regulators add an additional layer of specificity, arrest SNAREpin intermediates, lower the energy required for fusion, and couple membrane fusion to triggering signals. The functional activity of some of these regulators determines the plasticity of regulated exocytosis. (Part of a Multi-author Review)     

Membrane permeability changes duringRana oocyte maturation

A transition from an open system to a closed one must occur during the complex process of meiotic maturation of the amphibian oocyte. Membrane permeability to urea inRana oocytes following progesterone stimulation was determined, and the largest decrease was found to coincide with germinal vesicle breakdown. These findings suggest that the timing of the disappearance of membrane permeability correlates with developmental events that prepare the oocyte for a hostile environment.     

Membrane traffic in the secretory pathway

It is generally thought that microtubule-associated motors insure long-range movements of the secretory vesicles from the center of the cell to its periphery, while myosins insure short-range movements at the cell periphery. However, several of the myosins that have been reported during the last decade to be involved in the exocytic pathway are not processive, meaning that they do not have the ability to move cargos along actin polymers. We will review here the possible mechanisms by which these myosins could contribute to the traffic of secretory proteins from the Golgi complex to the plasma membrane.     

Factors affecting high-frequency fungal protoplast fusion

Summary Factors influencing the fusion frequency of protoplasts have been examined with auxotrophic mutants ofAspergillus nidulans. The optimum conditions were a total of 5 to 15 million protoplasts per ml, and 25% polyethylene glycol (PEG) 4000 or 6000 as fusogenic agent in 10 to 100 mM CaCl₂ solution.