The SALMFamides: a new family of neuropeptides isolated from an echinoderm.

We have isolated two novel related neuropeptides from the radial nerve cords of the starfishes Asterias rubens and Asterias forbesi. One is an octapeptide with the amino acid sequence Gly-Phe-Asn-Ser-Ala-Leu-Met-Phe-NH2 and the other is a dodecapeptide with the amino acid sequence Ser-Gly-Pro-Tyr-Ser-Phe-Asn-Ser-Gly-Leu-Thr-Phe-NH2. The peptides were purified using high performance liquid chromatography (HPLC) and a radioimmunoassay for the molluscan FMRFamide-related neuropeptide, pQDPFLRFamide. Both peptides share minimal sequence identity with members of the family of FMRFamide-like peptides so we have designated them as founder members of a new family, the SALMFamides. We refer to the octapeptide as SALMFamide 1 (S1) and the dodecapeptide as SALMFamide 2 (S2). S1 and S2 are the first neuropeptides identified in species belonging to the phylum Echinodermata.     

Schistosome/mollusk: genetic compatibility

Schistosomiasis remains one of the most prevalent parasitic infections and has significant economic and public health consequences in many developing countries. Economic development and improvement in standard of living in these countries are dependent on the elimination of this odious disease. For the control of Schistosomiasis, understanding the host/parasite association is important, since the host parasite relationship is often complex and since questions remain concerning the susceptibility of snails to infection by respective trematodes and their specificity and suitability as hosts for continued parasite development. Thus, the long term aim of this research is to learn more about the genetic basis of the snail/parasite relationship with the hope of finding novel ways to disrupt the transmission of this disease. In the current research, genetic variability among susceptible and resistant strains within and between Biomphalaria glabrata and B. tenagophila was investigated using RAPD-PCR. The results indicate great genetic variations within the two snail species using three different primers (intrapopulational variations), while specimens from the same snail species showed few individual differences between the susceptible and resistant strains (interpopulational variation).     

Locustatachykinin I and II, two novel insect neuropeptides with homology to peptides of the vertebrate tachykinin family

Two myotropic peptides termed locustatachykinin I (Gly-Pro-Ser-Gly-Phe-Tyr-Gly-Val-Arg-NH2) and locustatachykinin II (Ala-Pro-Leu-Ser-Gly-Phe-Tyr-Gly-Val-Arg-NH2) were isolated from brain-corpora cardiaca-corpora allata-suboesophageal ganglion extracts of the locust, Locusta migratoria. Both peptides exhibit sequence homologies with the vertebrate tachykinins. Sequence homology is greater with the fish and amphibian tachykinins (up to 45%) than with the mammalian tachykinins. In addition, the intestinal myotropic activity of the locustatachykinins is analogous to that of vertebrate tachykinins. The peptides discovered in this study may just be the first in a whole series of substances from arthropod species to be identified as tachykinin family peptides. Moreover, both chemical and biological similarities of vertebrate and insect tachykinins substantiate the evidence for a long evolutionary history of the tachykinin peptide family.     

Innate immunity: involvement of new neuropeptides

Secretory granules of chromaffin cells from the adrenal medulla store catecholamines and a variety of peptides that are secreted in the extracellular medium during exocytosis. Among these fragments, several natural peptides displaying antimicrobial activities at the micromolar range have been isolated and characterized. We have shown that these peptides, derived from the natural processing of chromogranins (CGs), proenkephalin-A (PEA) and free ubiquitin (Ub), are released into the circulation and display antibacterial and antifungal activities. In this review we focus on three naturally secreted antimicrobial peptides corresponding to CGA1–76 (vasostatin-I), the bisphosphorylated form of PEA209–237 (enkelytin) and Ub. In addition, the antimicrobial properties of the synthetic active domains of vasostatin-I (CGA47–66 or chromofungin) and Ub (Ub65–76 or ubifungin) are reported.     

Discovery of a new prokaryotic type I GTP cyclohydrolase family

GTP cyclohydrolase I (GCYH-I) is the first enzyme of the de novo tetrahydrofolate biosynthetic pathway present in bacteria, fungi, and plants, and encoded in Escherichia coli by the folE gene. It is also the first enzyme of the biopterin (BH4) pathway in Homo sapiens, where it is encoded by a homologous folE gene. A homology-based search of GCYH-I orthologs in all sequenced bacteria revealed a group of microbes, including several clinically important pathogens, that encoded all of the enzymes of the tetrahydrofolate biosynthesis pathway but GCYH-I, suggesting that an alternate family was present in these organisms. A prediction based on phylogenetic occurrence and physical clustering identified the COG1469 family as a potential candidate for this missing enzyme family. The GCYH-I activity of COG1469 family proteins from a variety of sources (Thermotoga maritima, Bacillus subtilis, Acinetobacter baylyi, and Neisseria gonorrhoeae) was experimentally verified in vivo and/or in vitro. Although there is no detectable sequence homology with the canonical GCYH-I, protein fold recognition based on sequence profiles, secondary structure, and solvation potential information suggests that, like GCYH-I proteins, COG1469 proteins are members of the tunnel-fold (T-fold) structural superfamily. This new GCYH-I family is found in approximately 20% of sequenced bacteria and is prevalent in Archaea, but the family is to this date absent in Eukarya.     

Crystal structure of Staphylococcus aureus Zn-glyoxalase I: new subfamily of glyoxalase I family

The crystal structures of protein SA0856 from Staphylococcus aureus in its apo-form and in complex with a Zn²⁺-ion have been presented. The 152 amino acid protein consists of two similar domains with α + β topology. In both crystalline state and in solution, the protein forms a dimer with monomers related by a twofold pseudo-symmetry rotation axis. A sequence homology search identified the protein as a member of the structural family Glyoxalase I. We have shown that the enzyme possesses glyoxalase I activity in the presence of Zn²⁺, Mg²⁺, Ni²⁺, and Co²⁺, in this order of preference. Sequence and structure comparisons revealed that human glyoxalase I should be assigned to a subfamily A, while S. aureus glyoxalase I represents a new subfamily B, which includes also proteins from other bacteria. Both subfamilies have a similar protein chain fold but rather diverse sequences. The active sites of human and staphylococcus glyoxalases I are also different: the former contains one Zn-ion per chain; the latter incorporates two of these ions. In the active site of SA0856, the first Zn-ion is well coordinated by His58, Glu60 from basic molecule and Glu40*, His44* from adjacent symmetry-related molecule. The second Zn3-ion is coordinated only by residue His143 from protein molecule and one acetate ion. We suggest that only single Zn1-ion plays the role of catalytic center. The newly found differences between the two subfamilies could guide the design of new drugs against S. aureus, an important pathogenic micro-organism.     

A new bioactive compound from the roots of Petasites hybridus

A new benzofuran derivative (1) was isolated from the roots of Petasites hybridus and its structure was determined as 1-(6-hydroxy-2-isopropenyl-1-benzofuran-5-yl)-1-ethanone by spectroscopic data and X-ray crystallographic analysis. The compound 1 showed moderate inhibitory activity on human breast cancer MCF-7 cells proliferation in vitro with an IC50 value of 48 μmol/L. The interaction of bovine serum albumin (BSA) and DNA with compound 1 were studied in an aqueous solution under physiological conditions by UV–vis spectroscopy.     

Diversification of the insulin receptor family in the helminth parasite Schistosoma mansoni

Insulin signalling is a very ancient and well conserved pathway in metazoan cells, dependent on insulin receptors (IR) which are transmembrane proteins with tyrosine kinase activity. A unique IR is usually present in invertebrates whereas two IR members are found with different functions in vertebrates. This work demonstrates the existence of two distinct IR homologs (SmIR-1 and SmIR-2) in the parasite trematode Schistosoma mansoni. These two receptors display differences in several structural motifs essential for signalling and are differentially expressed in parasite tissues, suggesting that they could have distinct functions. The gene organization of SmIR-1 and SmIR-2 is similar to that of the human IR and to that of the IR homolog from Echinococcus multilocularis (EmIR), another parasitic platyhelminth. SmIR-1 and SmIR-2 were shown to interact with human pro-insulin but not with pro-insulin-like growth factor-1 in two-hybrid assays. Phylogenetic results indicated that SmIR-2 and EmIR might be functional orthologs whereas SmIR-1 would have emerged to fulfil specific functions in schistosomes.     

Spatial organization and conformational peculiarities of the callatostatin family of neuropeptides.

The structures and conformational peculiarities of five members of the callatostatin family of neuropeptides, i.e. Leu- and Met-callatostatins, ranging in size from 8 to 16 amino acid residues have been investigated by a theoretical conformational analysis method. A comparative analysis of the conformational flexibilities of Met-callatostatin with those of the hydroxylated analogues, [Hyp2]- and [Hyp3]-Met-callatostatin has been carried out. Helically packed C-terminal pentapeptide in the structure of all investigated Leu-callatostatins are shown to be possible. The reason for the great number low-energy conformers for the callatostatin N-terminus is discussed.     

A new non-HLA multigene family associated with thePERB11 family within theMHC class I region

 In an effort to initiate steps designed to characterize the idiopathic hemochromatosis disease gene, the HLA-A/HLA-F region where this gene is in disequilibrium linkage with some polymorphic markers has been overlapped by a yeast artificial chromosome (YAC) contig. In order to achieve the physical mapping of these YACs and of the corresponding genomic region, we subcloned one of the YACs involved. A computer-assisted analysis of the sequence of one subclone led to the isolation of a potential exon that proved to belong to a new expressed messenger named HCGIX. After Southern blot analysis, the corresponding cDNA clone was found to belong to a new multigene family whose members are dispersed throughout the HLA class I region and are closely associated with members of another recently described multigene family designated PERB11. The data reported here suggest that these two multigene families form a cluster that have been dispersed together throughout the telomeric part of the major histocompatibility complex and have been involved in the genesis of this human class I region. Received: 23 February 1996 / Revised: 23 April 1996     

Flight substrates and their regulation by a member of the AKH/RPCH family of neuropeptides in Cerambycidae

The pattern of metabolic changes during tethered flight with lift-generation was investigated in two South African species of long-horned beetles (family: Cerambycidae), namely Phryneta spinator and Ceroplesis thunbergi. Energy substrates were measured in haemolymph and flight muscles at rest, after a flight period of 1 min at an ambient temperature of 25-29 degrees C, and 1 h thereafter. Flight diminished the levels of proline and carbohydrates in the haemolymph and proline and glycogen in the flight muscles of both species, and caused an increase in the levels of alanine in both compartments. The concentration of lipids in the haemolymph, however, was not changed upon flight in either species. The resting period of 1 h following a 1 min flight episode, was sufficient to reverse the metabolic situation in haemolymph and flight muscles to pre-flight levels in both species. Purification of an extract of the corpora cardiaca from the two beetle species on RP-HPLC, resulted in the isolation and subsequently in the identification (by mass spectrometry, Edman degradation and RP-HPLC) of an octapeptide of the AKH/RPCH family, denoted Pea-CAH-I (pGlu-Val-Asn-Phe-Ser-Pro-Asn-Trpamide), present in each species. It was demonstrated that low doses of Pea-CAH-I elicited increases in the concentration of proline, as well as carbohydrates, in the haemolymph of both species. The levels of lipids, however, remained unchanged upon injection of this peptide. It is concluded that, upon stimulation by flight, the peptide Pea-CAH-I is released from the corpus cardiacum of a cerambycid beetle and is responsible for the regulation of the major flight substrates, proline and carbohydrates, of these beetles.     

The spatial organization and conformational flexibility of neuropeptides of the gallatostatin family

The spatial organization and conformational flexibility of neuropeptides of the gallatostatin family was studied by the method of theoretical conformational analysis. It was found that the spatial organization of neuropeptides allows the realization of folded helical structures of the C-terminal pentapeptide, and the flexibility of neuropeptides is due to a great number of low-energy states in the N-terminal fragment of the molecule.     

CRISPR/Cas9: A new tool for the study and control of helminth parasites

Recent reports of CRISPR/Cas9 genome editing in parasitic helminths open up new avenues for research on these dangerous pathogens. However, the complex morphology and life cycles inherent to these parasites present obstacles for the efficient application of CRISPR/Cas9‐targeted mutagenesis. This is especially true with the trematode flukes where only modest levels of gene mutation efficiency have been achieved. Current major challenges in the application of CRISPR/Cas9 for study of parasitic worms thus lie in enhancing gene mutation efficiency and overcoming issues involved in host passage so that mutated parasites survive. Strategies developed for CRISPR/Cas9 studies on Caenorhabditis elegans, protozoa and mammalian cells, including novel delivery methods, the choice of selectable markers, and refining mutation precision represent novel tactics whereby these impediments can be overcome. Furthermore, employing CRISPR/Cas9‐mediated gene drive to interfere with vector transmission represents a novel approach for the control of parasitic worms that is worthy of further exploration.     

Nuocytes and beyond: new insights into helminth expulsion

T helper 2 (Th2) responses, characterized by the expression of the type-2 cytokines interleukin (IL)-4, IL-5, IL-9 and IL-13, are essential for the control of parasitic helminth infections and also drive the pathogenesis of allergy and asthma. Such responses are initiated, maintained and regulated, in part, by an array of innate effector cells and cytokines. However, relatively little is known about how the initiation of type-2 immune responses occurs in vivo. The recent discovery, using helminth models, of several novel innate immune cells capable of shaping type-2 immune responses allows us to reflect on the progress made in this area. It also affords us the opportunity to highlight the diversity of immune responses that can be driven by innate cells responding rapidly to early cytokine cues.