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1.
Protein Pept Lett ; 24(7): 668-675, 2017.
Article in English | MEDLINE | ID: mdl-28462721

ABSTRACT

BACKGROUND: Sleep is a natural part of every individual's life. Delta sleep-inducing peptide (DSIP) is a nonapeptide that could promote sleep through the induction of slow wave sleep. However, little is known about the pharmacological effect of DSIP on insomnia. OBJECTIVES: The main objective of this study was to analyze the pharmacological effect of DSIP on insomnia. METHODS: We designed a fusion protein containing N-terminal TAT-based transduction domain followed by human serum albumin and DSIP and designated this protein as PHD fusion protein. The PHD fusion protein were expressed in Pichia pastoris and purified. Mice were administered single subcutaneous injections three concentrations of PHD fusion protein (0.5, 1, 2 mg/kg), and the pharmacological activity of PHD fusion protein was studied using classic pentobarbitalinduced sleep test. RESULTS: We expressed the PHD fusion protein in P. pastoris; furthermore, the PHD fused protein was purified to near homogeneity by DEAE Sepharose FF, Phenyl Sepharose HP and Blue Sepharose 6 FF. Our result showed that the increase of pentobarbital-induced hypnotic effect characterized by reducing sleep latency and prolonged sleep duration was observed for increasing concentrations of PHD fusion protein (P<0.05); moreover, different dose of PHD fusion protein could induce the mice to re-sleep in a dose-dependent manner, whereas higher doses of PHD fusion protein (1.0, 2.0 mg/kg) significantly increased the rate of sleep re-onset compared with the vehicle group of mice (P<0.05). CONCLUSION: PHD fusion protein increased the hypnotic effects of pentobarbital by reducing sleep latency and prolonged sleep duration. The present study suggested PHD fusion protein could be a new drug candidate for insomnia.


Subject(s)
Delta Sleep-Inducing Peptide/administration & dosage , Recombinant Fusion Proteins/administration & dosage , Sleep Initiation and Maintenance Disorders/drug therapy , Sleep/drug effects , Animals , Delta Sleep-Inducing Peptide/chemistry , Delta Sleep-Inducing Peptide/genetics , Humans , Mice , Pentobarbital/administration & dosage , Pichia/genetics , Protein Domains , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Serum Albumin/administration & dosage , Serum Albumin/chemistry , Serum Albumin/genetics , Sleep Initiation and Maintenance Disorders/pathology
2.
Mater Sci Eng C Mater Biol Appl ; 42: 461-5, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25063142

ABSTRACT

Various biomolecules, for example proteins, peptides etc., entrapped in polymer matrices, impact interactions between matrix and cells, including stimulation of cell adhesion and proliferation. Delta-sleep inducing peptide (DSIP) possesses numerous beneficial properties, including its abilities in burn treatment and neuronal protection. DSIP entrapment in two macroporous polymer matrices based on copolymer of dimethylaminoethyl methacrylate and methylen-bis-acrylamide (Co-DMAEMA-MBAA) and copolymer of acrylic acid and methylen-bis-acrylamide (Co-AA-MBAA) has been studied. Quite 100% of DSIP has been entrapped into positively charged Co-DMAEMA-MBAA matrix, while the quantity of DSIP adsorbed on negatively charged Co-AA-MBAA was only 2-6%. DSIP release from Co-DMAEMA-MBAA was observed in saline solutions (0.9% NaCl and PBS) while there was no DSIP release in water or 25% ethanol, thus ionic strength was a reason of this process.


Subject(s)
Acrylamides/chemistry , Delta Sleep-Inducing Peptide/isolation & purification , Delta Sleep-Inducing Peptide/pharmacokinetics , Methacrylates/chemistry , Polymers/chemistry , Adsorption , Delta Sleep-Inducing Peptide/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate , Hydrogen-Ion Concentration , Materials Testing , Porosity , Sodium Chloride
3.
Biomed Khim ; 59(1): 65-75, 2013.
Article in Russian | MEDLINE | ID: mdl-23650723

ABSTRACT

The aim of the study was to entrap delta-sleep inducing peptide (DSIP) in cross-linked poly(vinyl alcohol)-based hydrogels of different structures and to evaluate peptide release kinetics from these hydrogels using an in vitro model. Isotropic and macroporous hydrogels on the basis of poly(vinyl alcohol) acrylic derivative (Acr-PVA) as well as macroporous hydogels containing epoxy groups which were synthesized by copolymerization of this monomer with glycidyl methacrylate. The isotropic hydrogels were fabricated at positive temperatures while the macroporous hydrogels (cryogels) were prepared at the temperatures below zero. The peptide was entrapped into macroporous modified PVA hydrogels by addition of a peptide solution on previously fabricated matrices, while into PVA-GMA hydrogels containing epoxy groups peptide immobilization was carried out by incubation of hydrogel matrices in the peptide solution. In the case of isotropic hydrogels the peptide was added into the polymer mixture at a hydrogel formation reaction. The peptide release kinetics was studied by incubation of hydrogels in PBS (pH 7.4), in physiological solution (0.9% NaCl) and in water. DSIP concentration in supernatants was determined by phase-reverse HPLC. DSIP release from the macroporous PVA hydrogel after 30 min incubation was 74, 70 and 64% in water, PBS and 0.9% NaCl, relatively, and it was completed in 3 hs. From the isotropic hydrogel the release neither peptide nor products of its degradation was not observed even after 48 hs of incubation. For freshly prepared hydrogel the release kinetics was as follows: 27 and 78% in 30 and 33 hs, relatively. In the case of the lyophilized hydrogel samples the peptide release was 63% in 30 min incubation while drying patterns at room temperature for 3 days resulted in significant peptide loss because its structure damage.


Subject(s)
Delta Sleep-Inducing Peptide/chemistry , Hydrogels/chemistry , Immobilized Proteins/chemistry , Models, Chemical , Polyvinyl Alcohol/chemistry , Delayed-Action Preparations , Humans , Kinetics
4.
Bioorg Khim ; 39(3): 277-84, 2013.
Article in Russian | MEDLINE | ID: mdl-24397026

ABSTRACT

We have undertaken a comparative study on physiological activity of well known neuropeptide DSIP (WAGGDASG E) and new closely related peptide KND (WKGGNASGE) in vivo assays. The sequence of K2, N5-DSIP (KND) was found recently by the computer search for DSIP homologous sequences in available nucleotide and protein databases at 324-332 site of Lysine-specific demethylase 3 B (EC 1.14.11, Swiss-Prot: Q7LBC6.1, 1-1761aa). This human lysine-specific histone demethylase is a representative of the recently discovered family of so called JmjC-domain-containing histone demethylases encoded by JMJD1B gene and ubiquitously expressed in tissues of various mammalian species. Biological investigations performed in this work confirm our preliminary data that DSIP-related peptide KND exhibits the similar biological properties in comparison with DSIP. Assessed by us antioxidative, anticonvulsive and behavioral effects of KND were even more expressed than in DSIP case. These results provide the additional evidences to support our suggestion that KND can be a possible endogenous prototype of "real" DSIP.


Subject(s)
Behavior, Animal/drug effects , Delta Sleep-Inducing Peptide/administration & dosage , Jumonji Domain-Containing Histone Demethylases/metabolism , Peptides/administration & dosage , Altitude , Animals , Antioxidants/administration & dosage , Delta Sleep-Inducing Peptide/chemistry , Humans , Hypoxia/drug therapy , Hypoxia/pathology , Jumonji Domain-Containing Histone Demethylases/genetics , Lipid Peroxidation/drug effects , Pentylenetetrazole/toxicity , Peptides/chemistry , Peptides/metabolism , Rats , Seizures/chemically induced , Seizures/drug therapy , Seizures/pathology
5.
Anal Chem ; 84(2): 1056-62, 2012 Jan 17.
Article in English | MEDLINE | ID: mdl-22132761

ABSTRACT

Arising from spontaneous aspartic acid (Asp) isomerization or asparagine (Asn) deamidation, isoaspartic acid (isoAsp, isoD, or beta-Asp) is a ubiquitous nonenzymatic modification of proteins and peptides. Because there is no mass difference between isoaspartyl and aspartyl species, sensitive and specific detection of isoAsp, particularly in complex samples, remains challenging. Here we report a novel assay for Asp isomerization by isotopic labeling with (18)O via a two-step process: the isoAsp peptide is first specifically methylated by protein isoaspartate methyltransferase (PIMT, EC 2.1.1.77) to the corresponding methyl ester, which is subsequently hydrolyzed in (18)O-water to regenerate isoAsp. The specific replacement of (16)O with (18)O at isoAsp leads to a mass shift of 2 Da, which can be automatically and unambiguously recognized using standard mass spectrometry, such as collision-induced dissociation (CID), and data analysis algorithms. Detection and site identification of several isoAsp peptides in a monoclonal antibody and the ß-delta sleep-inducing peptide (DSIP) are demonstrated.


Subject(s)
Delta Sleep-Inducing Peptide/chemistry , Isoaspartic Acid/analysis , Mass Spectrometry , Oxygen Radioisotopes , Protein D-Aspartate-L-Isoaspartate Methyltransferase/metabolism , Amino Acid Sequence , Chromatography, High Pressure Liquid , Molecular Sequence Data
6.
Peptides ; 32(4): 826-31, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21262293

ABSTRACT

Delta sleep inducing peptide (WAGGDASGE, DSIP) is a well known multifunctional regulatory peptide. Numerous studies have confirmed its stress-protective and adaptive activity which is independent of the origin or nature of the stress or other harmful factors. However, the biosynthetic origin of DSIP remains obscure, since nothing is known of its protein precursor(s) and their encoding gene(s). We have performed a comprehensive analysis of available gene and protein databases for homologous peptide sites within mammalian resources including man. A family of Jumonji C (JmjC)-domain-containing histone demethylases was shown to contain a sequence fragment closely homologous to DSIP. One type of these ubiquitous and phylogenetically ancient proteins encoded by JMJD1B gene includes the WKGGNASGE sequence that differs from DSIP by only 2 amino acid residues in positions 2 and 5. The respective peptide was synthesized and its biological effects were evaluated in a preliminary way in the forced swimming and antitoxic tests. We suggest that the histone demethylases of the JmjC-group containing DSIP-related region can be considered as possible protein precursors of endogenous peptides with DSIP-like activity.


Subject(s)
Delta Sleep-Inducing Peptide/metabolism , Jumonji Domain-Containing Histone Demethylases/metabolism , Amino Acid Sequence , Animals , Delta Sleep-Inducing Peptide/chemistry , Humans , Jumonji Domain-Containing Histone Demethylases/chemistry , Male , Molecular Sequence Data , Rats
7.
J Neurochem ; 97(2): 303-9, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16539679

ABSTRACT

Delta sleep-inducing peptide (DSIP) was isolated from rabbit cerebral venous blood by Schoenenberger-Monnier group from Basel in 1977 and initially regarded as a candidate sleep-promoting factor. However, the link between DSIP and sleep has never been further characterized, in part because of the lack of isolation of the DSIP gene, protein and possible related receptor. Thus the hypothesis regarding DSIP as a sleep factor is extremely poorly documented and still weak. Although DSIP itself presented a focus of study for a number of researchers, its natural occurrence and biological activity still remains obscure. DSIP structure is different from any other known representative of the various peptide families. In this mini-review we hypothesize the existence of a DSIP-like peptide(s) that is responsible (at least partly) for DSIP-like immunoreactivity and DSIP biological activity. This assumption is based on: (i) a highly specific distribution of DSIP-like immunoreactivity in the neurosecretory hypothalamic nuclei of various vertebrate species that are not particularly relevant for sleep regulation, as revealed by the histochemical studies of the Geneva group (Charnay et al.); (ii) a large spectrum of DSIP biological activity revealed by biochemical and physiological studies in vitro; (iii) significant slow-wave sleep (SWS) promoting activity of certain artificial DSIP structural analogues (but not DSIP itself!) in rabbits and rats revealed by our early studies; and (iv) significant SWS-promoting activity of a naturally occurring dermorphin-decapeptide that is structurally similar to DSIP (in five of the nine positions) and the sleep-suppressing effect of its optical isomer, as revealed in rabbits. Potential future studies are outlined, including natural synthesis and release of this DSIP-like peptide and its role in neuroendocrine regulation.


Subject(s)
Delta Sleep-Inducing Peptide/physiology , Sleep/physiology , Animals , Delta Sleep-Inducing Peptide/chemistry , Delta Sleep-Inducing Peptide/metabolism , Humans
9.
Izv Akad Nauk Ser Biol ; (4): 467-74, 2001.
Article in Russian | MEDLINE | ID: mdl-11525128

ABSTRACT

The sleep-inducing activity of Delta Sleep-Inducing Peptide (DSIP) and its 13 synthetic analogs has been studied on rabbits with preliminary implanted electrodes. The peptides were injected into the lateral ventricle of cerebrum. Polygraphic computer monitoring of sleep-wake states was carried out at daytime for 7-12 h. DSIP and most analogs had no statistically significant effect on sleep compared to the control administration of saline to the same animals. [NMeAla2]DSIP and [Pro2]DSIP had a pronounced sleep-inducing effect and reliably increased the proportion of slow-wave sleep by 10-15% on average compared to the control. Several other analogs had a week sleep-inducing effect, increasing the proportion of slow-wave sleep during specific recording time only. [beta-Ala2]DSIP significantly suppressed sleep. In addition, this analog, as well as parent DSIP and four proline-containing nonapeptides, slightly increased the body temperature. The revealed differences may be due to both conformation properties and proteolytic resistance of the studied molecules, and it may reflect their indirect involvement in the control sleep-wake hormonal processes.


Subject(s)
Delta Sleep-Inducing Peptide/analogs & derivatives , Delta Sleep-Inducing Peptide/pharmacology , Sleep/drug effects , Animals , Delta Sleep-Inducing Peptide/chemistry , Electroencephalography , Male , Peptides/chemistry , Peptides/pharmacology , Rabbits , Sleep/physiology , Structure-Activity Relationship
10.
Arch Biochem Biophys ; 349(2): 225-35, 1998 Jan 15.
Article in English | MEDLINE | ID: mdl-9448709

ABSTRACT

Static and dynamic spectroscopic properties of the tryptophanil emission in conjunction with circular dichroism (CD) spectroscopy and molecular dynamics are used to investigate the interactions of the neuropeptide neuromedin B (NMB) and the membrane-permeable delta sleep-inducing peptide (DSIP) with the membrane lipid phase. Our data indicate that in solution both peptides exist in energetically equivalent conformations, whereas in the presence of the membrane specific conformational states are stabilized. By changing from the aqueous to the lipid phase, the static and the dynamic fluorescence properties of the NMB's tryptophan residue are clearly affected: the fluorescence steady-state spectrum as well as the resolved fluorescence decay-associated spectra (DAS) are shifted to the blue with a significant increase of the fluorescence intensity of the second lifetime component (tau 2-DAS). On the other hand, in the lipid environment the same parameters of DSIP are negligibly affected as compared to the aqueous buffer. The CD and molecular dynamics analyses are consistent with these results and indicate that, while NMB assumes a helix-like conformation with the tryptophan residue in the apolar surface, DSIP adopts a globule-like structure with the indole ring that is surface-exposed. As previously found for neuromedin C (Polverini, E., Neyroz, P., Fariselli, P., Casadio, R., and Masotti, L., Biochem. Biophys. Res. Commun. 214, 663-668, 1995), for NMB the stabilized "lipophilic" structure also may favor the correct peptide-receptor contact and recognition. For DSIP, the lipid-stabilized conformation does not support an amphiphilic structure-driven peptide-membrane interaction and suggests a hydrophobicity-driven diffusion across the bilayer.


Subject(s)
Delta Sleep-Inducing Peptide/chemistry , Lipid Bilayers , Neurokinin B/analogs & derivatives , Protein Conformation , Circular Dichroism , Dimyristoylphosphatidylcholine , Fluorescence Polarization , Lysophosphatidylcholines , Models, Chemical , Models, Molecular , Neurokinin B/chemistry , Phosphatidylserines , Spectrometry, Fluorescence/methods
11.
J Biol Chem ; 272(49): 30918-27, 1997 Dec 05.
Article in English | MEDLINE | ID: mdl-9388238

ABSTRACT

The 77-residue delta sleep-inducing peptide immunoreactive peptide (DIP) is a close homolog of the Drosophila melanogaster shortsighted gene product. Porcine DIP (pDIP) and a peptide containing a leucine zipper-related partial sequence of pDIP, pDIP(9-46), was synthesized and studied by circular dichroism and nuclear magnetic resonance spectroscopy in combination with molecular dynamics calculations. Ultracentrifugation, size exclusion chromatography, and model calculations indicated that pDIP forms a dimer. This was confirmed by the observation of concentration-dependent thermal folding-unfolding transitions. From CD spectroscopy and thermal folding-unfolding transitions of pDIP(9-46), it was concluded that the dimerization of pDIP is a result of interaction between helical structures localized in the leucine zipper motif. The three-dimensional structure of the protein was determined with a modified simulated annealing protocol using experimental data derived from nuclear magnetic resonance spectra and a modeling approach based on an established strategy for coiled coil structures. The left-handed super helical structure of the leucine zipper type sequence resulting from the modeling approach is in agreement with known leucine zipper structures. In addition to the hydrophobic interactions between the amino acids at the heptade positions a and d, the structure of pDIP is stabilized by the formation of interhelical i to i' + 5 salt bridges. This result was confirmed by the pH dependence of the thermal-folding transitions. In addition to the amphipatic helix of the leucine zipper, a second helix is formed in the NH2-terminal part of pDIP. This helix exhibits more 310-helix character and is less stable than the leucine zipper helix. For the COOH-terminal region of pDIP no elements of regular secondary structure were observed.


Subject(s)
Delta Sleep-Inducing Peptide/chemistry , Leucine Zippers , Neuropeptides/chemistry , Repressor Proteins , Acetylation , Amino Acid Sequence , Animals , Binding Sites , DNA/metabolism , Drosophila melanogaster , Humans , Models, Molecular , Molecular Sequence Data , Protein Denaturation , Protein Structure, Secondary , Sequence Alignment , Solutions , Static Electricity , Swine , Transcription Factors/chemistry
12.
Biochim Biophys Acta ; 1309(3): 200-4, 1996 Dec 11.
Article in English | MEDLINE | ID: mdl-8982256

ABSTRACT

We have cloned a 420 bp cDNA from a human fetal brain cDNA library in lambda encoding the human homologue of a DSIP-immunoreactive leucine zipper protein (DIP) isolated from porcine brain. The derived human protein (hDIP) shares a significant sequence identity with parts of the murine TSC-22 and Drosophila shs, both proteins which are discussed as functioning as transcriptional regulators. A similar role of hDIP is partially confirmed by the results of an RT-PCR analysis, demonstrating the widespread distribution of the protein among different human tissues.


Subject(s)
Delta Sleep-Inducing Peptide/chemistry , Leucine Zippers/genetics , Proteins/chemistry , Repressor Proteins , Transcription, Genetic/genetics , Amino Acid Sequence , Animals , Base Sequence , Blotting, Southern , Brain/embryology , Cloning, Molecular , Conserved Sequence , DNA, Complementary/chemistry , Drosophila/metabolism , Gene Expression Regulation/genetics , Humans , Mice , Molecular Sequence Data , Sequence Analysis , Sequence Homology, Amino Acid
13.
Biochem Mol Biol Int ; 38(4): 739-45, 1996 Apr.
Article in English | MEDLINE | ID: mdl-8728103

ABSTRACT

The spatial structure of cyclo(-Gly-DSIP-), a physiologically active analog of the delta sleep-inducing peptide, was determined by computer modelling using 1H NMR data. An interesting feature of the spatial structure in DMSO was detected. One side of almost planar resulting conformation is formed by the side chains of the Asp5, Ser7 and Glu9 residues, the side chain of the Trp1 residue forming the other part of the outer surface. This feature may be associated with the functional properties of the peptide.


Subject(s)
Delta Sleep-Inducing Peptide/analogs & derivatives , Amino Acid Sequence , Chemical Phenomena , Chemistry, Physical , Computer Simulation , Delta Sleep-Inducing Peptide/chemistry , Magnetic Resonance Spectroscopy , Molecular Sequence Data , Protein Conformation
14.
Peptides ; 16(6): 1153-6, 1995.
Article in English | MEDLINE | ID: mdl-8532601

ABSTRACT

We measured morning plasma concentrations of delta sleep-inducing-peptide-like-immunoreactivity (DSIP-LI) in 9 sleep apnea patients, 10 narcolepsy patients, and 11 normal controls. Comparisons between the three groups showed no significant differences, although there was a trend toward association with low levels of DSIP-LI in the narcoleptic group, particularly in patients not using medications. No differences were found in the morning or evening plasma DSIP-LI levels in a second group of 11 normal controls and 8 sleep apneics. Our findings do not appear to support a biological marker role of disease activity for single measures of plasma DSIP in sleep apnea.


Subject(s)
Delta Sleep-Inducing Peptide/blood , Narcolepsy/blood , Sleep Apnea Syndromes/blood , Adult , Amino Acid Sequence , Biomarkers/blood , Case-Control Studies , Delta Sleep-Inducing Peptide/chemistry , Female , Humans , Immunohistochemistry , Male , Middle Aged , Molecular Sequence Data , Narcolepsy/physiopathology , Sleep Apnea Syndromes/physiopathology , Sleep Stages/physiology
15.
Biochemistry ; 33(6): 1323-31, 1994 Feb 15.
Article in English | MEDLINE | ID: mdl-8312250

ABSTRACT

Peptides and peptide-like molecules as a class have very poor permeability through biological membranes, which severely compromises their potential effectiveness as therapeutic agents. In order to gain insight into the problem of delivering peptide and protein drugs and to establish a model in which the effects of systematic structural variations on transport can be explored, an investigation of the solution conformation of a membrane-permeable peptide was undertaken. Delta-sleep-inducing peptide (DSIP, MW 849) was used in this investigation. DSIP is a charged, hydrophilic peptide that possesses the unusual ability to diffuse passively across the blood-brain barrier (BBB) in vivo [Kastin, A. J., Banks, W. A., Castellanos, P. F., Nissen, C., & Coy, D. H. (1982) Pharmacol. Biochem. Behav. 17, 1187-1191] and across monolayers of brain microvessel endothelial cells in vitro, a model of the BBB [Raeissi, S., & Audus, K. L. (1989) J. Pharm. Pharmacol. 41, 848-852]. This nonapeptide was studied in solution using one- and two-dimensional nuclear magnetic resonance (NMR), circular dichroism (CD), Fourier transform infrared (FT-IR), and fluorescence spectroscopies in conjunction with molecular modeling. Our spectroscopic findings suggest that DSIP exists in a dynamic equilibrium between unordered and folded structures. Residues 2-5 and 6-9 tend to form type I beta-turns in aqueous solution and a similar, but more ordered, helix-like structure inducible in 40% trifluoroethanol (TFE). NMR, FT-IR, and CD studies in aqueous solution support the dynamic equilibrium hypothesis with the IR data, suggesting that the beta-turn population is approximately 40%.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Cell Membrane Permeability , Delta Sleep-Inducing Peptide/chemistry , Amino Acid Sequence , Circular Dichroism , Computer Simulation , Delta Sleep-Inducing Peptide/metabolism , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Sequence Data , Protein Conformation , Protein Folding , Protein Structure, Secondary , Solutions , Spectrometry, Fluorescence , Spectroscopy, Fourier Transform Infrared , Thermodynamics
16.
Biol Signals ; 1(2): 78-87, 1992.
Article in English | MEDLINE | ID: mdl-1307916

ABSTRACT

We have recently demonstrated that delta-sleep-inducing peptide (DSIP) stimulates indolamine secretion from rat pineal glands. In the present study, we show that tryptophan (TRP), as well as DSIP, stimulate melatonin (MEL) and 5-methoxy-tryptophol (5-ML) secretion in a dose-dependent manner between 5 x 10(-6) and 10(-4) M. The kinetic characteristics of the MEL and 5-ML secretion and the response induced by the two substances were similar. The increase in MEL secretion in response to 10(-4) M DSIP was completely inhibited by pretreatment of the pineals with 10(-5) M phenanthroline (amino-peptidase inhibitor), suggesting that stimulatory effect of DSIP was due to TRP liberated by peptide degradation. This mechanism occurring in the pineal was confirmed using 10(-4) M para-chlorophenylalamine (TRP hydroxylase inhibitor), which reduced the pineal response to 10(-4) and 10(-5) M DSIP by 50 and 100%, respectively.


Subject(s)
Delta Sleep-Inducing Peptide/pharmacology , Pineal Gland/drug effects , Tryptophan/pharmacology , Amino Acid Sequence , Animals , Delta Sleep-Inducing Peptide/chemistry , Fenclonine/pharmacology , In Vitro Techniques , Indoles/metabolism , Kinetics , Male , Melatonin/metabolism , Molecular Sequence Data , Perfusion , Phenanthrolines/pharmacology , Pineal Gland/metabolism , Rats , Rats, Wistar , Tryptophan/metabolism
17.
C R Acad Sci III ; 314(6): 259-62, 1992.
Article in French | MEDLINE | ID: mdl-1318772

ABSTRACT

The "delta sleep inducing peptide" (DSIP) is a regulatory peptide localized in the brain, the hypophysis and some endocrine cells of the gut. The present immunological study, performed with a monoclonal antibody to DSIP, provides evidence for the presence of DSIP-like immunoreactivity (DSIP-LI) in a strain of small cell carcinoma. The specificity of the immunoreaction was assessed by the tests using heterologous antigen known to be secreted by these cells. The DSIP could play a role in the course of this disease.


Subject(s)
Carcinoma, Small Cell/metabolism , Delta Sleep-Inducing Peptide/chemistry , Lung Neoplasms/metabolism , Carcinoma, Small Cell/pathology , Humans , Immunohistochemistry , Lung Neoplasms/pathology , Tumor Cells, Cultured/chemistry
18.
Peptides ; 12(6): 1375-7, 1991.
Article in English | MEDLINE | ID: mdl-1815223

ABSTRACT

A phosphorylated analogue of DSIP at Ser7 has been shown to exist endogenously by immunochemical studies. An enzyme which could phosphorylate DSIP has not yet been identified. In the present study, we examined DSIP as a substrate for in vitro phosphorylation by casein kinase II. DSIP was phosphorylated by the enzyme with apparent Km and Vmax values of 20 mM and 90.9 nmol/min/mg protein, respectively. Both ATP and GTP were utilized as phosphoryl donors. Phosphorylation of DSIP was inhibited by heparin and enhanced by spermine. These results demonstrate that DSIP can serve as a possible substrate for casein kinase II in vitro.


Subject(s)
Delta Sleep-Inducing Peptide/metabolism , Protein Kinases/metabolism , Adenosine Triphosphate/metabolism , Amino Acid Sequence , Animals , Casein Kinases , Delta Sleep-Inducing Peptide/chemistry , Guanosine Triphosphate/metabolism , In Vitro Techniques , Kinetics , Molecular Sequence Data , Oligopeptides/chemistry , Oligopeptides/metabolism , Phosphorylation , Rabbits , Substrate Specificity
19.
Peptides ; 12(3): 445-54, 1991.
Article in English | MEDLINE | ID: mdl-1923924

ABSTRACT

The naturally occurring forms of delta sleep-inducing peptide (DSIP) are not fully identified. In the present study, porcine pituitaries and adrenal glands were extracted in water, saline or acid under various conditions and immunoreactive DSIP (IR-DSIP) quantified by radioimmunoassay. The highest concentrations were measured in anterior pituitary extracts (40.8 +/- 2.6 ng/g tissue weight) recovered using water with aprotinin. However, high performance liquid chromatography (HPLC) indicated degradation of hydrophobic forms of IR-DSIP in water extracts. Extraction in acetic acid including C18 Sep-Pak purification resulted in an elution profile of IR-DSIP in adrenal extracts with a major peak coeluting with synthetic DSIP [DSIP(1-9)], whereas anterior pituitary extract showed material of higher hydrophobicity. Approximately 30% of IR-DSIP in anterior pituitary as well as in adrenal gland extracts seemed to be glucosylated, as based on concanavalin A chromatography. One of the DSIP-immunoreactive components by immunoblotting (molecular mass 25 kDa) was identified in both pituitary and adrenal gland extracts. In conclusion, several chromatographically distinct forms of IR-DSIP are present in the porcine pituitary and adrenal gland. IR material eluting as DSIP(1-9) is present in adrenal gland extract. The procedure and solution used for tissue extraction seem to be essential in order to obtain reliable elution positions on HPLC.


Subject(s)
Adrenal Glands/chemistry , Delta Sleep-Inducing Peptide/isolation & purification , Pituitary Gland/chemistry , Amino Acid Sequence , Animals , Chromatography , Delta Sleep-Inducing Peptide/chemistry , Delta Sleep-Inducing Peptide/immunology , Immunoblotting , Immunochemistry , Molecular Sequence Data , Radioimmunoassay , Swine
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