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C Mollay

Publications and source records attributed to C Mollay.

18 recordsLinked to original sources
PubMed1999

Bv8, a small protein from frog skin and its homologue from snake venom induce hyperalgesia in rats.

From skin secretions of Bombina variegata and Bombina bombina, we isolated a small protein termed Bv8. The sequence of its 77 amino acids was established by peptide analysis and by cDNA cloning of the Bv8 precursor. Bv8 stimulates the contraction of the guinea-pig ileum at nanomolar concentrations. The contraction is not inhibited by a variety of antagonists. Injection of a few micrograms of Bv8 into the brain of rats elicits, as assessed by the tail-flick test and paw pressure threshold, a marked hyperalgesia which lasts for about 1 h. Bv8 is related to protein A, a component of the venom of the black mamba. After i.c.v. injection, protein A is even more active than Bv8 in inducing hyperalgesia.

Amino Acid Sequence↗
PubMed1996

BSTI, a trypsin inhibitor from skin secretions of Bombina bombina related to protease inhibitors of nematodes.

From skin secretions of the European frog Bombina bombina, a new peptide has been isolated that contains 60 amino acids, including 10 cysteine residues. Its sequence was determined by automated Edman degradation and confirmed by analysis of the cDNA encoding the precursor. A search in the databanks demonstrated that the pattern of cysteine residues in this skin peptide is similar to the ones found in protease inhibitors from Ascaris and in a segment of human von Willebrand factor. The 3D structure of the trypsin inhibitor from Ascaris suum could be used as a template to build a model of the amphibian peptide. In addition, we have demonstrated that this constituent of skin secretion is indeed an inhibitor of trypsin and thrombin, with K(i) values in the range of 0.1 to 1 microM. The new peptide was thus named BSTI for Bombina skin trypsin/thrombin inhibitor.

Amino Acid Sequence↗
PubMed1996

The lipocalin Xlcpl1 expressed in the neural plate of Xenopus laevis embryos is a secreted retinaldehyde binding protein.

The cellular and structural properties and binding capabilities of a lipocalin expressed in the early neural plate of Xenopus laevis embryos and the adult choroid plexus have been investigated. It was found that this lipocalin, termed Xlcpl1, binds retinal at a nanomolar concentration, retinoic acid in the micromolar range, but does not show binding to retinol. Furthermore, this protein also binds D/L thyroxine. The Xlcpl1 cDNA was expressed in cell culture using the vaccinia virus expression system. In AtT20 cells, Xlcpl1 was secreted via the constitutive secretory pathway. We therefore assume that cpl1 binds retinaldehyde during the transport through the compartments of the secretory pathway that are considered to be the storage compartments of retinoids. Therefore, cpl1-expressing cells will secrete the precursors of active retinoids such as retinoic acid isomers. These retinoids may enter the cytosol by diffusion or receptor-controlled mechanisms, as has been shown for exogenously applied retinoids. Based on these data, it is suggested that cpl1 is an integral member of the retinoid signaling pathway and, therefore, it plays a key role in pattern formation in early embryonic development.

Amino Acid Sequence↗
PubMed1992

Purification and properties of an iminopeptidase from culture media of Streptomyces plicatus.

The degradation of the prosequence of the secreted enzyme endo-beta-N-acetylglucosaminidase H from Streptomyces plicatus is not elucidated. Both the primary structure of this segment and the finding that the secreted species contain ragged aminoterminal ends of specific structure suggested that a dipeptidylaminopeptidase might mature this enzyme. Therefore, we tested the culture medium of Streptomyces plicatus for prolin-specific peptidases. Proline iminopeptidase was purified about 800-fold to homogeneity from the culture medium. Dipeptidylaminopeptidase, the enzyme that seemed most likely to process the prosequence of endo-beta-N-acetylglucosaminidase H, could not be detected.

Aminopeptidases↗
PubMed1989

Export of honeybee prepromelittin in Escherichia coli depends on the membrane potential but does not depend on proteins secA and secY.

Honeybee prepromelittin (70 amino acid residues), the precursor of an eukaryotic secretory protein, and a hybrid protein between prepromelittin and mouse dihydrofolate reductase (257 amino acid residues) were expressed in Escherichia coli and characterized with respect to their requirements for transport across the plasma membrane. Both precursor proteins are posttranslationally processed and exported into the periplasm, and they both depend on the membrane potential for this to occur. With respect to dependence on components of the export machinery, however, the two precursor proteins show striking differences: the small precursor protein prepromelittin does not require the function of proteins secA and secY; the large precursor protein prepromelittin-dihydrofolate reductase, on the other hand, depends on both components. The implications of these observations with respect to the mechanisms of protein export in E. coli and of protein import into the endoplasmic reticulum are discussed.

Animals↗
PubMed1986

Isolation of a dipeptidyl aminopeptidase, a putative processing enzyme, from skin secretion of Xenopus laevis.

A dipeptidyl aminopeptidase has been purified to apparent homogeneity from skin secretion of Xenopus laevis. This enzyme is a glycoprotein with a molecular mass of about 98 kDa. It hydrolyzes a variety of dipeptidyl-p-nitroanilides and oligopeptides containing proline, alanine or glycine as the second amino acid and is inhibited by diisopropylfluorophosphate. The pH optimum was found to be around 8, while at pH 6, substrates were cleaved at about one-third of the maximal rate. This dipeptidyl aminopeptidase has the specificity required for the cleavage of amino-terminal extensions preceding the sequence of caerulein and xenopsin in their respective precursors.

Animals↗
PubMed1986

Import of honeybee prepromelittin into the endoplasmic reticulum. Requirements for membrane insertion, processing, and sequestration.

Honeybee prepromelittin is correctly processed and imported by dog pancreas microsomes. Membrane insertion of prepromelittin, assayed as signal sequence removal by signal peptidase, is not dependent on signal recognition particle and docking protein. However, a previously uncharacterized proteinaceous component of the microsomal membrane is required for completion of membrane transfer of promelittin. Furthermore, membrane insertion of prepromelittin is not coupled to translation. These data suggest the signal sequence, in addition to its role in membrane recognition, has a more general function for membrane insertion, cotranslational import of proteins is not an intrinsic feature of microsomes, and at least in certain cases, proteinaceous membrane components are involved in membrane transfer.

Amino Acid Sequence↗
PubMed1986

Skin peptides in Xenopus laevis: morphological requirements for precursor processing in developing and regenerating granular skin glands.

The biosynthesis of the peptides caerulein and PGLa in granular skin glands of Xenopus laevis proceeds through a pathway that involves discrete morphological rearrangements of the entire secretory compartment. Immunocytochemical localization of these peptides during gland development indicates that biosynthetic precursors are synthesized in intact secretory cells, whereas posttranslational processing requires morphological reorganization to a vacuolated stage. The bulk of the processed secretory material is then stored in vacuolae-derived storage granules. In the mature gland, storage granules are still formed at a low level. However, in this case processing takes place in a distinct cytoplasmic structure, the multicored body, which we suggest to be functionally equivalent to vacuolae. When granular glands regenerate after having lost all their storage granules upon strong stimuli, another morphological pathway is used. 2 wk after gland depletion, secretory cells become arranged in a monolayer that covers the luminal surface of the gland. Storage granules are formed continuously within these intact secretory cells. Here, precursor processing does not require a vacuolated stage as in newly generated glands but occurs in multicored bodies. Most storage granules seem to be formed in the third week of regeneration. The high biosynthetic activity is also reflected by the high activity of the putative processing enzyme dipeptidyl aminopeptidase during this period of regeneration.

Animals↗
PubMed1983

Cooperative effects in the interaction between melittin and phosphatidylcholine model membranes. Studies by temperature scanning densitometry.

The interaction between the peptide melittin from bee venom with multilamellar liposomes of dipalmitoylphosphatidylcholine or egg yolk phosphatidylcholine has been studied by the method of temperature scanning densitometry, yielding information on the specific volume of the association products and on the changes during the thermotropic transition of the lipids. The effects of the interaction were found to be biphasic with respect to melittin concentration; below 10(-3) mol per mol of phospholipid, an increase in transition temperature, abolition of the pretransition, a reduction in the transition volume of the lipids by about 25%, and a nonadditive increase in apparent specific volume of the complexes were observed. Only minor changes in these parameters could be detected between molar ratios of 10(-3) and 10(-2). Above 1 mol % melittin, the transition temperature decreased and the transition volume approached zero around 10 mol %. Since the effects in the low concentration range cannot be accounted for only by local perturbations around the actual sites of interaction, it is concluded that the interaction involves long range effects of melittin affecting several hundreds of phospholipid molecules. The results are discussed in terms of a phospholipid cluster model, whereby the interaction with melittin leads to a cooperative transition of entire clusters to a state of expanded volume. It is suggested that this transition may be important to the activating effect of melittin on membrane enzymes and to enhanced membrane permeability and lysis.

Bee Venoms↗
PubMed1982

Comparative effects of melittin and its hydrophobic and hydrophilic fragments on bilayer organization by Raman spectroscopy.

For bilayer systems consisting of 1,2-dimyristoyl phosphatidylcholine (DMPC) incubated with melittin, a polypeptide capable of integrating itself within the membrane, temperature profiles derived from Raman spectroscopic data indicate the existence of an immobilized lipid annulus surrounding the polypeptide. In particular, temperature profiles derived from C--H, C--D and C--C stretching mode parameters for 25:1, 14:1 and 10:1 lipid:protein mole ratios exhibit two order-disorder transitions. The primary (lower) gel to liquid crystalline phase transition is depressed when polypeptide concentration is increased. The concentration-independent higher temperature transition is associated with a fluidization of the immobilized boundary lipids present at the lipid-polypeptide interface within the bilayer. We estimate that five to seven lipids are involved in this discrete boundary layer around the inserted membrane component. The behavior of the intrinsic hydrophobic (residues 1-19) and of the extrinsic hydrophilic (residues 20-26) portions of melittin in the bilayer is compared with the properties of the intact polypeptide. We emphasize evidence that both intrinsic and extrinsic components immobilize lipids contiguous to the polypeptide.

Animals↗
PubMed1982

Cleavage of honeybee prepromelittin by an endoprotease from rat liver microsomes: identification of intact signal peptide.

It has previously been shown that rat liver microsomes contain a proteolytic enzyme that cleaves honeybee prepromelittin to yield promelittin. This enzyme has now been further purified by centrifugation on a sucrose-deoxycholate gradient and then reconstituted into phospholipid vesicles. Incubation of prepromelittin with vesicles in the presence of melittin yields, in addition to promelittin, a hydrophobic peptide. The latter could be isolated by extraction with l-butanol and paper electrophoresis in 30% formic acid and was shown to be intact signal peptide by analysis of peptic fragments and automated Edman degradation. The microsomal enzyme is thus an endoprotease that hydrolyzes prepromelittin exclusively at the pre-pro junction. The precision of this cleavage of an insect preprotein by a rat liver enzyme indicates that we are dealing with the ubiquitous eukaryotic signal peptidase.

Amino Acid Sequence↗
PubMed1980

Interaction of melittin with dimyristoyl phosphatidylcholine liposomes: evidence for boundary lipid by Raman spectroscopy.

The interaction of melittin, a polypeptide consisting of 26 amino acid residues, with dimyristoyl phosphatidylcholine bilayers was investigated by vibrational Raman spectroscopy. Spectral peak height intensity ratios, involving vibrational transitions in both the 3000 cm-1 acyl chain methylene carbon-hydrogen stretching mode region and the 1100 cm-1 acyl chain carbon-carbon skeletal stretching mode interval, served as temperature profile indices for monitoring the bilayer order-disorder processes. For a lipid : melittin molar ratio of 14 : 1 two order-disorder transitions were observed. In comparison to a gel to liquid crystalline phase transition of 22.5 degrees C for the pure lipid, the lower transition, exhibiting a 2 degree C width, is centered at 17 degrees C and is associated with a depression of the main lipid phase transition of dimyristoyl phosphatidylcholine. The second thermal transition, displaying a 7 degree C interval, occurs at approx. 29 degrees C and is associated with the melting behavior of approximately seven immobilized boundary lipids which surround the inserted hydrophobic segment of the polypeptide. For a lipid : melittin molar ratio of 10 : 1 two thermal transitions are also observed at 11 and 30 degrees C. As before, they represent, respectively, the main gel to liquid crystalline phase transition and the melting behavior of approximately four boundary lipids attached to melittin. From these data alternative schemes are suggested for disposing the immobilized lipids around the hydrophobic portion of the polypeptide within the bilayer.

Bee Venoms↗
PubMed1976

Action of phospholipases on the cytoplasmic membrane of Escherichia coli. Stimulation by melittin.

The emission maximum of the single tryptophan residue of melittin was measured in the presence of phosphatidylethanolamine liposomes and Escherichia coli cytoplasmic membranes. In both cases, the fluorescence maximum was shifted to shorter wavelengths indicating a transfer of the indole ring to an apolar environment. E. coli membranes were labelled in position 2 of their phospholids with [14C]oleic acid. These membranes were used for measuring the activity of an endogenous phospholipase A2. A slow hydrolysis is observed, which can be accelerated by adding melittin. The extent of the stimulation depends on the molar ratio of melittin to membrane phospholipid. Under suitable conditions, the initial rate of hydrolysis is six to seven times higher in the presence than in the absence of melittin. The action of the phospholipase A2 from bee venom is also stimulated by melittin. An identical stimulation was observed with either E. coli membranes or pure phosphatidylethanolamine liposomes as substrate.

Cell Membrane↗