Mechanism of membrane damage mediated by human eosinophil cationic protein

Recent evidence suggests a role for eosinophil granule proteins in contact-dependent antibody-mediated cytotoxicity. Cytolysis may involve a secretory phenomenon whereby granule proteins are released at the site of contact between eosinophil and target cells. Several basic proteins have been isolated from eosinophil granules, including the major basic protein, eosinophil cationic protein, eosinophil protein-X and eosinophil peroxidase. One of the major granule proteins of human eosinophils is the eosinophil cationic protein (ECP) which has been shown to damage schistosomula of Schistosoma mansoni at concentrations as low as 10(-7). Here, we describe the formation of functional channels by purified human ECP. The transmembrane pores formed by ECP are relatively voltage-insensitive and non-ion-selective, suggesting a role for channel formation by ECP in target cell damage mediated by eosinophils. Channel formation by granule proteins of immune effector cells may represent a general and effective mechanism of target cell killing.     

Structural basis for modulation and agonist specificity of HCN pacemaker channels

The family of hyperpolarization-activated, cyclic nucleotide-modulated (HCN) channels are crucial for a range of electrical signalling, including cardiac and neuronal pacemaker activity, setting resting membrane electrical properties and dendritic integration. These nonselective cation channels, underlying the I(f), I(h) and I(q) currents of heart and nerve cells, are activated by membrane hyperpolarization and modulated by the binding of cyclic nucleotides such as cAMP and cGMP. The cAMP-mediated enhancement of channel activity is largely responsible for the increase in heart rate caused by beta-adrenergic agonists. Here we have investigated the mechanism underlying this modulation by studying a carboxy-terminal fragment of HCN2 containing the cyclic nucleotide-binding domain (CNBD) and the C-linker region that connects the CNBD to the pore. X-ray crystallographic structures of this C-terminal fragment bound to cAMP or cGMP, together with equilibrium sedimentation analysis, identify a tetramerization domain and the mechanism for cyclic nucleotide specificity, and suggest a model for ligand-dependent channel modulation. On the basis of amino acid sequence similarity to HCN channels, the cyclic nucleotide-gated, and eag- and KAT1-related families of channels are probably related to HCN channels in structure and mechanism.     

A collimated, high-speed outflow from the dying star V Hydrae

Stars with masses in the range 1-8 solar masses (M(\circ)) live ordinary lives for approximately 10(9)-10(10) years, but die extraordinary deaths. First, during their death throes as asymptotic giant branch (AGB) stars they eject, over 10(4)-10(5) years, half or more of their mass in slowly expanding, spherical winds, and then, in a short (a few 100-1,000 years) and poorly understood phase, they are transformed into aspherical planetary nebula. Recent studies support the idea that high-speed, jet-like flows play a crucial role in this transformation. Evidence for such outflows is indirect, however; this phase is so short that few nearby stars are likely to be caught in the act. Here we report the discovery of a newly launched, high-speed jet-like outflow in the nearby AGB star, V Hydrae. We have detected both proper motions and ongoing evolution in the jet. These results support a model in which the jet is driven by an accretion disk around an unseen, compact companion. We also find a central, dense equatorial disk-like structure which may enable and/or enhance the formation of the accretion disk.     

The causes of Charcot-Marie-Tooth disease

Charcot-Marie-Tooth (CMT) disease serves as the summary term for the most frequent forms of inherited peripheral neuropathies that affect motor and sensory nerves. In the last 12 years, 14 genes have been identified that cause different CMT subforms. The genes found initially are predominantly responsible for demyelinating and dysmyelinating neuropathies. Genes affected in axonal and rare forms of CMT have only recently been identified. In this review, we will focus on the currently known genes that are associated with CMT syndromes with regards to their genetics and function.Received 5 April 2003; received after revision 20 May 2003; accepted 23 May 2003     

Management Philosophy and Practices of Habitat Conservation for Jiuzhaigou Nature Reserve, Sichuan, China

Starting from the pressure on habitat of Jiuzhaigou Nature Reserve brought about by traveling activity, this paper puts forward such philosophy of community co-management of conservation-based tourism development for Jiuzhaigou Nature Reserve, According to this philosophy, emphasis is laid on the analysis of the mission of habitat protection, functional division, habitat maintenance, community construction, scientific study and management organization system. The study shows that the close relation and significance of rights and interests shared between nature reserve and resi-dents in community, as well as the value of community comanagement philosophy of conservation-based tourism development in sustainable management of nature reserve, have set an exemplary example for other nature reserves.     

Distal residues in the oxygen binding site of haemoglobin studied by protein engineering

The geometries of the Fe-O2 and Fe-CO bonds in myoglobin and haemoglobin differ significantly from those in free porphyrin model compounds. It has been suggested that steric hindrance by Val-E11 and His-E7 and a hydrogen bond between His-E7 and oxygen affect the geometry and electronic state of the Fe-ligand bond, and that these interactions may be important in controlling oxygen affinity. We have produced mutant haemoglobins in E. coli having Val(67 beta)E11 replaced by Ala, Met, Leu or Ile and His(58 beta)E7 by Gln, Val or Gly. We have studied the effect of these mutations on the equilibrium and kinetics of ligand binding. The conformation of the new side chains and their effect on the protein structure have been examined by X-ray crystallography, and the vibrational properties of the Fe-CO bond observed by resonance Raman spectroscopy. We found that the steric hindrance of ligand binding by the E11 residue and the polarity of the E7 residue in the beta subunit are critical for fine-tuning ligand affinity.