Molecular biological strategies in the analysis of antibiotic peptide gene families. The use oligonucleotides as hybridization probes.

Origins and Development of Peptide Antibiotic Research: From Extracts to Abstracts to Contracts, John K. Spitznagel. Part I. Isolation and Characterization of Antibacterial Peptides. HPLC Methods for Purification of Antimicrobial Peptides, Michael E. Selsted. Strategies for the Isolation and Characterization of Antimicrobial Peptides of Invertebrates, Charles Hetru and Philippe Bulet. Big Defensin and Tachylectins-1 and -2, Shun-ichiro Kawabata and Sadaaki Iwanaga. Tachyplesin and Anti-Lipopolysaccharide Factor, Tatsushi Muta and Sadaaki Iwanaga. Circular Dichroism Studies of Secondary Structure of Peptides, Martha M. Juban, Maryam M. Javadpour, and Mary D. Barkley. Analytical Ultracentrifugation Studies of Association of Peptides, Martha M. Juban, Maryam M. Javadpour, and Mary D. Barkley. NMR Characterization of Amphipathic Helical Peptides, Xiaotang Wang and Kathleen M. Morden. Part II. Molecular Biology of Antibacterial Peptides. Laboratory Production of Antimicrobial Peptides in Native Conformation, Erika V. Valore and Tomas Ganz. An Approach Combining Rapid cDNA Amplification and Chemical Synthesis for the Identification of Novel, Cathelicidin-Derived, Antimicrobial Peptides, Alessandro Tossi, Marco Scocchi, Margherita Zanetti, Renato Gennaro, Paola Storici, and Domenico Romeo. Molecular Biological Strategies in the Analysis of Antibiotic Peptide Gene Families: The Use of Oligonucleotides as Hybridization Probes, Charles L. Bevins and Gill Diamond. Part III. Assay Systems for Studying Antibacterial Peptides. Designer Assays for Antimicrobial Peptides: Disputing the 'One-Size-Fits-All' Theory, Deborah A. Steinberg and Robert I. Lehrer. Interaction of Cationic Peptides with Bacterial Membranes, Shafique Fidai, Susan W.Farmer, and Robert E. W. Hancock. The Genetic Basis of Microbial Resistance to Antimicrobial Peptides, Eduardo A. Groisman and Arden Aspedon. Assay of Antibacterial Activities of the Bactericidal/Permeability-Increasing Proeting in Natural Biological Fluids, Jerrold Weiss. Assay Systems for Measurement of Chemotactic Activity, H. Anne Pereira. Neutralization of the In Vivo Activity of E. coli-Derived Lipopolysaccharide by Cationic Peptides, Daniel J. Brackett, Megan R. Lerner, and H. Anne Pereira. Index.

We have used phage-displayed peptide libraries to identify novel ligands to the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp120. Screening of libraries of random 12-mers, 7-mers, and cyclic 9-mers produced two families of gp120 binding peptides. Members of a family with the prototype sequence RINNIPWSEAMM (peptide 12p1) inhibit the interaction between gp120 and both four-domain soluble CD4 (4dCD4) and monoclonal antibody (MAb) 17b, a neutralizing antibody that covers the chemokine receptor binding surface on gp120. Peptide 12p1 inhibits the interaction of 4dCD4 with gp120 from three different HIV strains, implying that it binds to a conserved site on gp120. Members of a second family of peptides, with the prototype sequence TSPYEDWQTYLM (peptide 12p2), bind more weakly to gp120. They do not detectably affect its interaction with 4dCD4, but they enhance its binding to MAb 17b. A common sequence motif in the two peptide families and cross-competition for gp120 binding suggest that they have overlapping contacts. Their divergent effects on the affinity of gp120 for MAb 17b may indicate that their binding stabilizes distinct conformational states of gp120. The functional properties of 12p1 suggest that it might be a useful lead for the development of inhibitors of HIV entry.

Evolution of the neuropeptide Y family: New genes by chromosome duplications in early vertebrates and in teleost fishes

Phylogeny of the Cholecystokinin/Gastrin Family

Corticotropin-releasing factor (CRF)-induced behaviors are modulated by intravenous administration of teneurin C-terminal associated peptide-1 (TCAP-1)

Functional and structural features of γ-zeathionins, a new class of sodium channel blockers

Cell-to-Cell Communication during Lateral Root Development

The tachykinin (TK) and tachykinin-related peptide (TKRP) family represent one of the largest peptide families in the animal kingdom and exert their actions via a subfamily of structurally related G-protein-coupled receptors. In this study, we have identified a novel TKRP receptor from the Octopus heart, oct-TKRPR. oct-TKRPR includes domains and motifs typical of G-protein-coupled receptors. Xenopus oocytes that expressed oct-TKRPR, like TK and TKRP receptors, elicited an induction of membrane chloride currents coupled to the inositol phosphate/calcium pathway in response to Octopus TKRPs (oct-TKRP I-VII) with moderate ligand selectivity. Substance P and Octopus salivary gland-specific TK, oct-TK-I, completely failed to activate oct-TKRPR, whereas a Substance P analog containing a C-terminal Arg-NH2 exhibited equipotent activation of oct-TKRPs. These functional analyses prove that oct-TKRPs, but not oct-TK-I, serve as endogenous functional ligands through oct-TKRPR, although both of the family peptides were identified in a single species, and the importance of C-terminal Arg-NH2 in the specific recognition of TKRPs by TKRPR is conserved through evolutionary lineages of Octopus. Southern blotting of RT-PCR products revealed that the oct-TKRPR mRNA was widely distributed in the central and peripheral nervous systems plus several peripheral tissues. These results suggest multiple physiologic functions of oct-TKRPs as neuropeptides both in the Octopus central nervous system and in peripheral tissues. This is the first report on functional discrimination between invertebrate TKRPs and salivary gland-specific TKs.

The tachykinin peptide family certainly represents one of the largest peptide families described in the animal organism. So far, more than 40 tachykinins have been isolated from invertebrate (insects, worms, and molluscs), protochordate, and vertebrate (skin, gastrointestinal tract, peripheral and central nervous system) tissues. Substance P (SP), first identified by bioassay as early as 1931 but sequenced only in 1971, several years after the elucidation of the structure of eledoisin from molluscan tissues and of physalaemin from amphibian skin, may be considered as a prototype of the tachykinins. Hitherto, as many as 19 tachykinins have been isolated from amphibian integument, and eight additional peptides have been isolated from amphibian gut and brain. Counterparts of skin tachykinins in mammalian tissues are SP, neurokinin A, and neurokinin B. Three main receptor subtypes for the tachykinins have been identified (NK1, NK2, and NK3), but their number is probably destined to increase. It is obvious that the peripheral and central effects of the tachykinins may substantially vary depending on the activation of different receptor subtypes. Matters are further complicated by the frequent capacity of the single tachykinins to bind, although with different affinity, to more receptors. It has been recognized that tachykinins have a variety of effects in physiological and pathological conditions, and there is evidence suggesting intrinsic neuroprotective and neurodegenerative properties of these neuropeptides. This review provides an update on the current body of knowledge regarding tachykinin occurrence and distribution in the animal kingdom, from the lowest invertebrates to man, and the physiological and pharmacological actions of tachykinins outlining the pregnant importance of this large peptide family.

Members of the Sir2 family of NAD-dependent protein deacetylases regulate diverse cellular processes including aging, gene silencing, and cellular differentiation. Here, we report that the distant mammalian Sir2 homolog SIRT6 is a broadly expressed, predominantly nuclear protein. Northern analysis of embryonic samples and multiple adult tissues revealed mouse SIRT6 (mSIRT6) mRNA peaks at day E11, persisting into adulthood in all eight tissues examined. At the protein level, mSIRT6 was readily detectable in the same eight tissue types, with the highest levels in muscle, brain, and heart. Subcellular localization studies using both C- and N-terminal green fluorescent protein fusion proteins showed mSIRT6-green fluorescent protein to be a predominantly nuclear protein. Indirect immunofluorescence using antibodies to two different mSIRT6 epitopes confirmed that endogenous mSIRT6 is also largely nuclear. Consistent with previous findings, we did not observe any NAD+-dependent protein deacetylase activity in preparations of mSIRT6. However, purified recombinant mSIRT6 did catalyze the robust transfer of radiolabel from [32P]NAD to mSIRT6. Two highly conserved residues within the catalytic core of the protein were required for this reaction. This reaction is most likely mono-ADP-ribosylation because only the modified form of the protein was recognized by an antibody specific to mono-ADP-ribose. Surprisingly, we observed that the catalytic mechanism of this reaction is intra-molecular, with individual molecules of mSIRT6 directing their own modification. These results provide the first characterization of a Sir2 protein from phylogenetic class IV.

Evolution and phylogeny of the corticotropin-releasing factor (CRF) family of peptides: Expansion and specialization in the vertebrates

Peptide Signaling in Plant Development

Relationships between the FMRFamide-related peptides and other peptide families.

Many neuropeptides are members of peptide families, with multiple structurally similar isoforms frequently found even within a single species. This raises the question of whether the individual peptides serve common or distinct functions. In the accompanying paper, we found high isoform specificity in the responses of the lobster (Homarus americanus) cardiac neuromuscular system to members of the pyrokinin peptide family: only one of five crustacean isoforms showed any bioactivity in the cardiac system. Because previous studies in other species had found little isoform specificity in pyrokinin actions, we examined the effects of the same five crustacean pyrokinins on the lobster stomatogastric nervous system (STNS). In contrast to our findings in the cardiac system, the effects of the five pyrokinin isoforms on the STNS were indistinguishable: they all activated or enhanced the gastric mill motor pattern, but did not alter the pyloric pattern. These results, in combination with those from the cardiac ganglion, suggest that members of a peptide family in the same species can be both isoform specific and highly promiscuous in their modulatory capacity. The mechanisms that underlie these differences in specificity have not yet been elucidated; one possible explanation, which has yet to be tested, is the presence and differential distribution of multiple receptors for members of this peptide family.

Neuropeptides are perhaps the most diverse intercellular signaling molecules in the animal kingdom. Owing to the high-throughput methodologies using the high-performance liquid chromatography as well as the mass spectrometry, the number of peptides identified from both vertebrates and invertebrates are increasing. In many cases, the multiple structurally related peptides which are unequivocally considered to be members of the same peptide family are found in the nervous system. We call such peptides the multiple family peptides. Here, we show the structure, distribution, and action of two kinds of multiple family peptides recently identified in a marine mollusc, Aplysia.