Peptides (v.25, #9)

Peptide pheromones: an overview by Miriam Altstein (1373-1376).

Enterococcal peptide sex pheromones: synthesis and control of biological activity by Josephine R. Chandler; Gary M. Dunny (1377-1388).
The enterococcal pheromone-inducible plasmids such as pCF10 represent a unique class of mobile genetic elements whose transfer functions are induced by peptide sex pheromones. These pheromones are excreted by potential recipient cells and detected by plasmid-containing donor cells at the cell surface, where the pheromone is imported and signals induction of the plasmid transfer system. Pheromone is processed from a chromosomally encoded lipoprotein and excreted by both the donor and recipient cells, but a carefully controlled detection system prevents a response to self-pheromone while still allowing an extremely sensitive response to exogenous pheromone.
Keywords: pCF10; pAD1; pPD1; Enterococcus faecalis; Pheromone; Pheromone-inducible conjugative plasmid; PrgZ; PrgY; iCF10;

Peptide signaling in Staphylococcus aureus and other Gram-positive bacteria by Gholson J. Lyon; Richard P. Novick (1389-1403).
There are two basic types of bacterial communication systems—those in which the signal is directed solely at other organisms and those in which the signal is sensed by the producing organism as well. The former are involved primarily in conjugation; the latter in adaptation to the environment. Gram-positive bacteria use small peptides for both types of signaling, whereas Gram-negative bacteria use homoserine lactones. Since adaptation signals are autoinducers the response is population-density-dependent and has been referred to as “quorum-sensing”. Gram-negative bacteria internalize the signals which act upon an intracellular receptor, whereas Gram-positive bacteria use them as ligands for the extracellular receptor of a two-component signaling module. In both cases, the signal activates a complex adaptation response involving many genes.
Keywords: Peptide signaling; Staphylococcus aureus; Gram-positive bacteria; Histidine kinase;

Lantibiotics are produced by a variety of Gram-positive bacteria. The production of these peptides appears to be regulated at the transcriptional level in a cell-density-dependent manner in various bacteria. This phenomenon has been studied in detail for the production of nisin by Lactococcus lactis, and the production of the structurally similar subtilin by Bacillus subtilis. In this paper, the molecular mechanism underlying regulation of nisin and subtilin production is reviewed. This quorum sensing, autoregulatory module includes the lantibiotics themselves as peptide pheromones, the signal transduction by the corresponding two-component regulatory systems, and the lantibiotic-responsive promoter elements in the biosynthesis gene clusters. Finally, the exploitation of these regulatory characteristics for the development of highly effective controlled gene expression systems in Gram-positive bacteria is discussed.
Keywords: Antimicrobial peptide; Lantibiotic; Signal transduction; Peptide pheromone; Regulation; Quorum sensing;

Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters by Michiel Kleerebezem; Roger Bongers; Ger Rutten; Willem M. de Vos; Oscar P. Kuipers (1415-1424).
The production of the type I antimicrobial peptide (AMP) subtilin by Bacillus subtilis is regulated in a cell-density-dependent manner [Kleerebezem M, de Vos WM, Kuipers OP. The lantibiotics nisin and subtilin act as extracellular regulators of their own biosynthesis. In: Dunny GM, Winans SC, editors. Cell–cell signaling in bacteria. Washington, D.C., USA: ASM Press; 1999. p. 159–74; Stein T, Borchert S, Kiesau P, Heinzmann S, Kloss S, Klein C, Helfrich M, Entian KD. Dual control of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol 2002;44:403–16; Stein T, Heinzmann S, Kiesau P, Himmel B, Entian KD. The spa-box for transcriptional activation of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol 2003;47:1627–36]. Three subtilin-responsive promoter elements within the spaBTCSIFEGRK are controlled by the specific cis-acting sequence element called the spa-box, which represents the binding site of the subtilin regulator SpaR [Stein T, Heinzmann S, Kiesau P, Himmel B, Entian KD. The spa-box for transcriptional activation of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol 2003;47:1627–36]. Here, we describe the functional characterization of the spaB, spaS and spaI promoters by transcriptional fusion with a promoterless β-glucuronidase encoding gusA gene. Within these gusA fusion constructs, transcription initiation start sites of the spaS and spaI promoters were mapped to be located downstream of the spa-box, which is in contrast to previous reports [Banerjee S, Hansen JN. Structure and expression of a gene encoding the precursor of subtilin, a small protein antibiotic. J Biol Chem 1988;263:9508–14; Stein T, Heinzmann S, Kiesau P, Himmel B, Entian KD. The spa-box for transcriptional activation of subtilin biosynthesis and immunity in Bacillus subtilis. Mol Microbiol 2003;47:1627–36]. Nevertheless, all spa-promoters displayed typical cell-density-dependent activity in a subtilin-producing strain B. subtilis ATCC6633. Moreover, analysis of β-glucuronidase activities in a spaB mutant of B. subtilis ATCC6633 and a derivative of strain 168 that harbors the spaRK genes integrated in the chromosomal amyE locus, confirmed that these promoters are activated by subtilin-triggered, SpaRK-mediated, quorum-sensing control. Quantitative analysis showed that the spaS promoter strength at a given subtilin concentration appeared to be approximately five-fold higher than the spaB promoter, which in turn is approximately two-fold higher than the spaI promoter. Finally, it is shown that the elementary components involved in subtilin-mediated regulation are the two-component system, SpaRK, and a spa-box containing promoter.
Keywords: Bacillus subtilis; Subtilin; Peptide pheromone; Regulation; Quorum sensing;

Quorum sensing (QS) in Gram-negative bacteria is generally assumed to be mediated by N-acyl-homoserine lactone molecules while Gram-positive bacteria make use of signaling peptides. We analyzed the occurrence in Gram-negative bacteria of peptides and transporters that are involved in quorum sensing in Gram-positive bacteria. Many class II bacteriocins and inducing factors produced by lactic acid bacteria (LAB) and competence stimulating peptides (CSPs) synthesized by streptococci are processed by their cognate ABC-transporters during their secretion. During transport, a conserved leader sequence, termed the double-glycine motif (GG-motif), is cleaved off by the N-terminal domain of the transporter, which belongs to the Peptidase C39 protein family. Several peptides containing a GG-motif were recently described in Gram-negative bacteria (Trends Microbiol 2001;9:164–8). To screen for additional putative GG-motif containing peptides, an in silico strategy based on MEME, HMMER2.2 and Wise2 was designed. Using a curated training set, a motif model of the leader peptide was built and used to screen over 120 fully sequenced bacterial genomes. The screening methodology was applied at the nucleotide level as probably many small peptide genes have not been annotated and may be absent from the non-redundant databases. It was found that 33% of the screened genomes of Gram-negative bacteria contained one or more transporters carrying a Peptidase C39 domain, compared to 44% of the genomes of Gram-positive bacteria. The transporters can be subdivided into four classes on the basis of their domain organization. Genes coding for putative peptides containing 23–142 amino acids and a GG-motif were found in close association with genes coding for Peptidase C39 domain containing proteins. These peptides show structural similarity to bacteriocins and peptide pheromones of Gram-positive bacteria. The possibility of signal transduction based on peptide signaling in Gram-negative bacteria is discussed.
Keywords: Quorum sensing; Peptide pheromone; Signal transduction; Membrane transport; Double-glycine motif; Peptidase C39 domain; ABC-transporters; Protein secretion; Genome-wide screening; HSL; Autoinducer; Bacteriocin;

Mating in Saccharomyces cerevisiae is initiated by the secretion of diffusible peptide pheromones that are recognized by G protein-coupled receptors (GPCR). This review summarizes the use of the α-factor (WHWLQLKPGQPMY) – GPCR (Ste2p) interaction as a paradigm to understand the recognition between medium-sized peptide hormones and their cognate receptors. Studies over the past 15 years have indicated that the α-factor is bent around the center of the pheromone and that residues near the amine terminus play a central role in triggering signal transduction. The bend in the center appears not to be rigid and this flexibility is likely necessary for conformational changes that occur as the receptor switches from the inactive to active state. The results of synthetic, biological, biochemical, molecular biological, and biophysical analyses have led to a preliminary model for the structure of the peptide bound to its receptor. Antagonists for Ste2p have changes near the N-terminus of α-factor, and mutated forms of Ste2p were discovered that appear to favor binding of these antagonists relative to agonists. Many features of this yeast recognition system are relevant to and have counterparts in mammalian cells.
Keywords: Saccharomyces cerevisiae; Antagonists; G protein-coupled receptors; Mating pheromone;

The intracellular signal transduction pathway by which the yeast Saccharomyces cerevisiae responds to the presence of peptide mating pheromone in its surroundings is one of the best understood signaling pathways in eukaryotes, yet continues to generate new surprises and insights. In this review, we take a brief walk down the pathway, focusing on how the signal is transmitted from the cell-surface receptor-coupled G protein, via a MAP kinase cascade, to the nucleus.
Keywords: Yeast mating pheromone; Mitogen-activated protein kinase; Signal transduction; Saccharomyces cerevisiae;

The study of insect seminal fluid proteins provides a unique window upon adaptive evolution in action. The seminal fluid of Drosophila melanogaster contains over 80 proteins and peptides, which are transferred together with sperm by mating males. The functions of many of these substances are not yet known. However, those that have been characterized have marked effects on the reproductive success of males and females. For example, seminal fluid proteins and peptides can decrease female receptivity, can increase egg production and can increase sperm storage, and are necessary for sperm transfer and success in sperm competition. In this review we focus on the currently known functions of seminal fluid molecules and on new technologies and approaches that are enabling novel questions about their form and function to be addressed. We discuss how techniques for disrupting the production of seminal fluid proteins, such as homologous recombination and RNA interference, along with the use of microarrays and yeast two hybrid systems, should allow us to address ever more sophisticated questions about seminal fluid protein function. These and similar techniques promise to reveal the function of naturally-occurring variants of these proteins and hence the evolutionary significance of genetic variation for them.
Keywords: Drosophila melanogaster; Seminal fluid proteins; Accessory gland proteins; Acps; Mating; Fecundity; Receptivity; Sperm competition;

Sex pheromone biosynthesis in many moth species is controlled by a cerebral neuropeptide, termed pheromone biosynthesis activating neuropeptide (PBAN). PBAN is a 33 amino acid C-terminally amidated neuropeptide that is produced by neuroendocrine cells of the subesophageal ganglion (SEG). Studies of the regulation of sex pheromone biosynthesis in moths have revealed that this function can be elicited by additional neuropeptides all of which share the common C-terminal pentapeptide FXPRL-amide (X = S, T, G, V). In the past two decades extensive studies were carried out on the chemical, cellular and molecular aspects of PBAN and the other peptides (termed the pyrokinin (PK)/PBAN family) aiming to understand the mode of their action on sex pheromone biosynthesis. In the present review we focus on a few of these aspects, specifically on the: (i) structure–activity relationship (SAR) of the PK/PBAN family, (ii) characterization of the PK/PBAN receptor and (iii) development of a novel strategy for the generation of PK/PBAN antagonists and their employment in studying the mode of action of the PK/PBAN peptides.
Keywords: PBAN; PBAN-receptor; Moths; Sex pheromones; Insect neuropeptides; Neuropeptide antagonists; Rational design;

Crustacean peptide and peptide-like pheromones and kairomones by Dan Rittschof; Jonathan H. Cohen (1503-1516).
Crustacean peptide pheromones, kairomones, and substituted amino sugar kairomones are reviewed from a historical perspective. These crustacean information molecules are secondary functions of structural polymers. They are partial hydrolysis products, generated usually by the action of trypsin-like enzymes on proteins, and glycosidase enzymes on glycoproteins and proteoglycans. Structure–function studies based upon synthetic mimics of peptide information molecules show neutral amino acids with a basic carboxyl terminal are active in modifying physiological and or behavioral responses. Behaviorally active substituted amino sugar mimics are disaccharide hydrolysis products of heparin and chondroitin sulfate. Similar molecules are also used as information molecules by a variety of other marine organisms indicating they are a common biological theme.
Keywords: Crustacean; Kairomone; Pheromone; Information molecules; Behavior; Peptides; Glycoproteins; Proteoglycans; Substituted amino sugars;

Peptide pheromones in female Nereis succinea by Jörg D. Hardege; Helga Bartels-Hardege; Carsten T. Müller; Manfred Beckmann (1517-1522).
Female specimen of the ragworm, Nereis succinea, employs a tetra-peptide, cysteinyl–glutathione (CSSG) as mate recognition and gamete release pheromone during reproduction. In the present study we review the role of peptide-based pheromones in Nereid worms focusing on pheromone production in females. New results demonstrate that the female ragworms produce the pheromone in the course of the oocyte maturation directly correlated to the oocyte diameter. Nevertheless, CSSG production is significantly increasing during the reproductive process, the nuptial dance, itself. The concentrations of the pheromone, nereithione (CSSG) and those of the possible precursors, reduced glutathione (GSH) and oxidized glutathione (GSSG), in the coelomic fluid were determined by HPLC showing that during spawning the female worms excrete 68.5 ± 12.5 μg/h of the pheromone. GSH is mobilized, or synthesized and continuously converted into the pheromone enabling female N. succinea to maintain a constant level of pheromone release until the GSH reserves are depleted. Upon release CSSG induces not only gamete release in males but in doses as low as 10−8 to 10−9  M also significantly increases male swimming activity and as such may guide males towards the slower swimming females.
Keywords: Cysteine–glutathione disulfide; Nereid polychaetes; Pheromone production;

Peptide and protein pheromones in molluscs by Abraham J. Susswein; Gregg T. Nagle (1523-1530).
Pheromones have been implicated in the control of a number of behaviors in molluscs, but few peptide pheromones have been characterized in these animals. Peptide pheromones include: (1) a family of water-borne peptide pheromonal attractants (attractins) in the gastropod Aplysia that are released during egg laying and attract other Aplysia to form egg-laying and mating aggregations; (2) a tetrapeptide (ILME) in the cephalopod Sepia that elutes from egg masses and is thought to be involved in the transport of oocytes in the genital tract during egg laying; and (3) a Sepia sperm-attracting peptide (SepSAP; PIDPGVamide) that is released from oocytes during egg laying to facilitate external fertilization.
Keywords: Aplysia; Attractin; Bursatella; ILME; Loligo; Mollusc; Peptide pheromone; PIDPGVamide; Sepia;

Peptide pheromones in newts by Fumiyo Toyoda; Kazutoshi Yamamoto; Takeo Iwata; Itaru Hasunuma; Marco Cardinali; Gilberto Mosconi; Alberta M. Polzonetti-Magni; Sakae Kikuyama (1531-1536).
This article reviews the current state of understanding of reproductive pheromones in amphibians, focusing mainly on the purification and characterization of peptide pheromones in newts of the genus Cynops, molecular cloning of cDNAs encoding the pheromone molecules, and hormonal control of secretion of these pheromones. Pheromones that attract sexually developed female Cynops pyrrhogaster and C. ensicauda newts were isolated from the male abdominal glands. The C. pyrrhogaster and C. ensicauda pheromones are peptides, designated sodefrin and silefrin, with the amino acid sequences SIPSKDALLK and SILSKDAQLK, respectively. Each pheromone attracts only conspecific females. Molecular cloning of cDNAs encoding sodefrin and silefrin revealed the presence of precursor proteins that are considered to generate these pheromone peptides. Pheromone precursor mRNA levels and radioimmunoassayable pheromone concentrations in the abdominal glands were elevated by prolactin and androgen. Sexual dimorphism and hormone dependency of the responsiveness of vomeronasal epithelium to sodefrin were noted. Significance of pheromones in the form of peptide for those performing reproductive behavior in an aquatic environment was also discussed.
Keywords: Newts; Peptide pheromones; Prolactin; Androgen;

Peptides derived from the post-translational processing of preprosodefrin were isolated from an extract of the abdominal glands of male red-bellied newts Cynops pyrrhogaster obtained 5 months prior to the onset of the breeding season. Structural characterization of the peptides showed that the pheromone sodefrin (SIPSKDALLK) is stored in a biologically inactive COOH-terminally extended form (SIPSKDALLKISA). It follows, therefore, that the activation of a protease that cleaves at a Lys–Ile bond to generate the active pheromone must occur by the time of onset of reproductive behavior. Additional peptides (representing preprosodefrin-(146–175)-peptide and preprosodefrin-(159–173)-peptide), that are derived from the precursor by cleavage at monobasic and dibasic processing sites, were also purified from the extract. The isolation of paralogs of these peptides, including an inactive COOH-terminally extended form of [Asn10]sodefrin, provides evidence for the expression of multiple genes encoding preprosodefrin. PCR products derived from total RNAs from the abdominal gland of individual newts collected from three different regions of Japan were analyzed. The data confirm the existence of multiple genes encoding sodefrin and its variants whose expression varied according to the individuals and the regions. However, genes encoding sodefrin were found to be expressed in all the specimens sampled.
Keywords: Sodefrin; Pheromone; Post-translational processing; Amphibian;

Aphrodisin, an aphrodisiac lipocalin secreted in hamster vaginal secretions by Loïc Briand; Didier Trotier; Jean-Claude Pernollet (1545-1552).
Vertebrates communicate through pheromones, which favor biological regulations within each species. Aphrodisin, a protein belonging to the lipocalin superfamily, found in hamster vaginal secretions, is detected by the male accessory olfactory system and induces or facilitates its copulatory behavior. Although much is known about aphrodisin structure, the question of whether aphrodisin bears itself the pheromonal function or is simply a carrier for hydrophobic small pheromones has not been definitely solved. Arguments based on use of recombinant aphrodisin deprived of any natural ligand and its capability to convey hamster pheromonal compounds will be discussed, together with progresses concerning putative natural ligand(s).
Keywords: Aphrodisin; Bartholin's glands; Copulation; Lipocalins; Pheromone; Vaginal secretions;

Urinary proteins and the modulation of chemical scents in mice and rats by Robert J Beynon; Jane L Hurst (1553-1563).
The urine of mice, rats and some other rodents contains substantial quantities of proteins that are members of the lipocalin family. The proteins are thought to be responsible for the binding and release of low molecular weight pheromones, and there is now good evidence that they discharge this role, providing a slow release mechanism for volatile components of scent marks. However, the proteins may function as chemosignalling molecules in their own right, contributing one or more roles in the communication of individual identity and scent mark ownership. In this review, we summarize current understanding of the structure and function of these urinary proteins, and speculate about their role as supporters or as key participants in the elaboration of the complex chemosensory properties of a rodent scent mark.
Keywords: Major urinary protein; Mouse; Rat; MUP; Pheromone; Semiochemistry;

Oxytocin, vasopressin, and social recognition in mammals by Isadora F. Bielsky; Larry J. Young (1565-1574).
While pheromones may act as social memory signals, oxytocin and vasopressin acting in the brain appear to be critical for the neural processing of olfactory signatures used for social discrimination. Evidence from a variety of laboratories using a range of animal models, as well as an array of molecular and pharmacological techniques, have helped to determine the neuroanatomical and functional roles oxytocin and vasopressin play in social cognition. In this review we discuss the considerable evidence for the roles of oxytocin and vasopressin in social recognition in rats and mice, as well as in offspring recognition in sheep and mate preference in monogamous voles.
Keywords: Vasopressin; V1a receptor; Social recognition; Oxytocin; Oxytocin receptor; Neuropeptides;