| 8890190 |
Isolation, antimicrobial activities, and primary structures of hamster neutrophil defensins |
10.1128/iai.64.11.4444-4449.1996. |
Infect Immun |
Isolation, antimicrobial activities, and primary structures of hamster neutrophil defensins
Abstract
- Hamster (Mesocricetus auratus) neutrophil granules contain at least four microbicidal peptides belonging to the defensin family. These compounds were purified from granule acid extracts by reverse-phase chromatography and termed HaNP-1 to -4 (hamster neutrophil peptide). HaNP-1 and HaNP-3 revealed the most bactericidal activity, with a 50% inhibitory concentration of 0.3 to 0.8 microg/ml for Staphylococcus aureus and Streptococcus pyogenes strains. The HaNP-4 was always isolated in concentrations exceeding about 10 times the concentrations of other hamster peptides, but its antibacterial activity as well as that of HaNP-2 was relatively lower, probably as a result of conserved Arg residue substitutions. Other microorganisms were also tested, and generally, hamster defensins exhibited less potency against gram-negative bacteria. The amino acid sequence of hamster defensins showed a high percentage of identity to the sequence of mouse enteric defensins, reaching about 60% identical residues in the case of HaNP-3 and cryptdin 3.
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| 8890254 |
Protegrins: structural requirements for inactivating elementary bodies of Chlamydia trachomatis |
10.1128/iai.64.11.4863-4866.1996. |
Infect Immun |
Protegrins: structural requirements for inactivating elementary bodies of Chlamydia trachomatis
Abstract
- We tested 20 protegrins against Chlamydia trachomatis serovar L2 (L2/434/Bu). Five of the protegrins had native structures; the others included nonamidated, enantiomeric, and truncated variants and peptides with <2 disulfide bonds. Antichlamydial activity resided principally in residues 5 to 15 of native protegrin PG-1, and optimal activity required both intramolecular disulfide bonds.
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| 8890729 |
In-frame exon 2 deletion in insulin receptor RNA in a family with extreme insulin resistance in association with defective insulin binding: a case report |
10.1530/eje.0.1350357. |
Eur J Endocrinol |
In-frame exon 2 deletion in insulin receptor RNA in a family with extreme insulin resistance in association with defective insulin binding: a case report
Abstract
- The phenotype and allelic expression of the insulin receptor gene is presented in a family with a patient with type A insulin resistance. Compared to controls, insulin receptor binding in transformed lymphocytes was 100%. 33% and 13% in the father, mother and proband, respectively. Reduced insulin receptor binding co-segregated with altered insulin receptor mRNA expression; the mother and daughter expressed eight insulin receptor mRNA species, including a set of four normal sized and a set of four shorter mRNA transcripts. In the proband the levels of the normal sized mRNA transcripts were suppressed relative to the shorter transcripts. Reverse polymerase chain reaction (PCR) revealed that the shorter transcripts contained an in-frame deletion of exon 2. Sequencing of the entire insulin receptor coding region revealed a paternally inherited A to T substitution in nucleotide 3205, converting isoleucine 996 to phenylalanine, which does not co-segregate with reduced binding. Therefore, we hypothesize that two findings are necessary for the presentation of type A insulin resistance in this patient: an in-frame deletion of the insulin receptor exon 2 that codes for amino acids crucial for insulin binding: and an inhibition of expression of the paternal insulin receptor allele.
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| 8894806 |
Anti-allergic activity of cyclosporin-A metabolites and their interaction with the parent compound and FK 506 |
10.1016/0192-0561(96)84506-1. |
Int J Immunopharmacol |
Anti-allergic activity of cyclosporin-A metabolites and their interaction with the parent compound and FK 506
Abstract
- The ability of cyclosporin-A (CSA) and four of its metabolites M1, M17, M18 and M21, to inhibit antigen-stimulated release of beta-hexoseaminidase from IgE-sensitized rat basophilic leukemia cells (RBL-2H3), as an in vitro correlate of anti-allergic effect, was studied Metabolites M17, M1 and M21 were effective in inhibiting enzyme release, though less potent than the parent compound. The concentrations achieving 50% inhibition (IC50 values) were 53.3, 315.5 and 875.7 ng/ml for CSA, M17 and M1, respectively. M21 had approximately same IC50 as M1 while M18 was essentially inactive. At the highest concentration tested (1000 ng/ml) the mean maximum percentage inhibitions were 98.6, 79.5, 53.9, 48.6 and 12.2 for CSA, M17, M1, M21 and M18, respectively. The relative anti-allergic potency of the metabolites was similar to their reported relative immunosuppressive potency. Combinations of low concentrations of CSA and its metabolites were synergistic in inhibiting enzyme release whereas at higher concentrations interactions were either additive or antagonistic. Even the concentrations of the metabolites that have little or no activity when used alone also potentiated the effect of CSA. The immunosuppressor FK 506 was found to be about three times more potent than CSA in this system and the interactions between FK 506 (3, 10 and 30 ng/ml) and CSA (10, 30 and 100 ng/ml) or M17 (20, 100 and 500 ng/ml) were synergistic at all combinations. Both CSA and M17 synergized more strongly with FK 506 than they did between themselves. These results show that some metabolites of CSA, like the parent compound, possess anti-allergic effects and that at concentrations that are obtainable in transplant patients, synergistic interaction occurs between CSA and its metabolites, and this may be of some therapeutic significance.
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| 8900077 |
Annetocin, an annelid oxytocin-related peptide, induces egg-laying behavior in the earthworm, Eisenia foetida |
10.1002/(SICI)1097-010X(19961001)276:2<151::AID-JEZ8>3.0.CO;2-N. |
J Exp Zool |
Annetocin, an annelid oxytocin-related peptide, induces egg-laying behavior in the earthworm, Eisenia foetida
Abstract
- Annetocin, an oxytocin-related peptide which we isolated from the earthworm Eisenia foetida, induced a series of egg-laying-related behaviors in the earthworms. These stereotyped behaviors consisted of well-defined rotatory movements, characteristic body-shape changes, and mucous secretion from the clitellum. Each of these behaviors is known to be associated with formation of the cocoon in which eggs are deposited. In fact, some of the earthworms injected with annetocin (> 5 nmol) laid eggs. Such egg-laying-related behaviors except for oviposition were also induced by oxytocin, but not by Arg-vasopressin and some other bioactive peptides isolated from E. foetida. Furthermore, annetocin also induced these egg-laying-like behaviors in the leech Whitmania pigra, but not in the polychaete Perinereis vancaurica. These results suggest that annetocin plays some key role in triggering stereotyped egg-laying behaviors in terrestrial or fresh-water annelids that have the clitella.
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| 8901613 |
A tumor-selective somatostatin analog (TT-232) with strong in vitro and in vivo antitumor activity |
10.1073/pnas.93.22.12513. |
Proc Natl Acad Sci U S A |
A tumor-selective somatostatin analog (TT-232) with strong in vitro and in vivo antitumor activity
Abstract
- We report a series of new in vitro and in vivo data proving the selective antitumor activity of our somatostatin structural derivative, TT-232. In vitro, it inhibited the proliferation of 20 different human tumor cell lines in the range of 50-95% and induced a very strong apoptosis. In vivo TT-232 was effective on transplanted animal tumors (Colon 26, B16 melanoma, and S180 sarcoma) and on human tumor xenografts. Treatment of MDA-MB-231 human breast cancer xenografted in mice with low submaximal doses of TT-232 0.25 and 0.5 mg/kg of body weight (b.w.) caused an average 80% decrease in the tumor volume resulting in 30% tumor-free animals surviving for longer than 200 days. Treatment of prostate tumor (PC-3) xenografted animals with 20 mg/kg of b.w. of TT-232 for 3 weeks resulted in 60% decrease in tumor volume and 100% survival even after 60 days, while 80% of nontreated animals perished. We have demonstrated that TT-232 did not bind to the membrane preparation of rat pituitary and cortex and had no antisecretory activity. TT-232 was not toxic at a dose of 120 mg/kg of b.w. in mice. Long-term incubation (24 h) of tumor cells with TT-232 caused significant inhibition of tyrosine kinases in good correlation with the apoptosis-inducing effect. The level of p53 or KU86 did not change following TT-232 treatment, suggesting a p53-independent apoptotic effect. Preincubation of human breast cancer cells (MDA-MB-453) with TT-232 for 2 h decreased the growth factor receptor autophosphorylation. All of these data suggest that TT-232 is a promising and selective antitumor agent.
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| 8906634 |
Cyclo (LL-dimethionine) is not utilized by rats as a methionine source |
10.3177/jnsv.42.339. |
J Nutr Sci Vitaminol (Tokyo) |
Cyclo (LL-dimethionine) is not utilized by rats as a methionine source
Abstract
- A low casein diet supplemented with 0.2-0.3% L-methionine promotes growth in rats, although it causes liver fat accumulation. Oligo-L-methionine on the other hand, prepared by papain-catalyzed oligomerization of L-methionine ethylester, stimulates growth in rats without causing liver fat accumulation, when added to a 8% casein diet. Oligo-L-methionine is a mixture of oligomers, with a oligomerization degree of 5 to 12, and is hydrolyzed very slowly by pancreatic juice and everted gut rings of rats. A methionine derivative, cyclo (LL-dimethionine), was examined in this study as it is hydrolyzed and/or absorbed very slowly from the intestinal tract and it was expected to be utilized as a methionine source without causing liver fat accumulation, when added to a low casein diet. It consists of two methionine residues linked together by two cis-peptide bonds and, therefore, is expected to be hydrolyzed and/or absorbed very slowly from intestinal tract. However, although cyclo (LL-dimethionine) was absorbed slowly from the intestinal tract, it was not utilized by rats as a methionine source, when added to 8% casein.
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| 8913308 |
A consensus structure for omega-conotoxins with different selectivities for voltage-sensitive calcium channel subtypes: comparison of MVIIA, SVIB and SNX-202 |
10.1006/jmbi.1996.0576. |
J Mol Biol |
A consensus structure for omega-conotoxins with different selectivities for voltage-sensitive calcium channel subtypes: comparison of MVIIA, SVIB and SNX-202
Abstract
- The omega-conotoxins are a set of structurally related peptides that have a wide range of specificities for different subtypes of the voltage-sensitive calcium channel (VSCC). To understand their VSCC subtype differentiation we studied the structure of two naturally occurring omega-conotoxins, MVIIA (specific to N-type) and SVIB (specific to P/Q-type) and a synthetic hybrid, SNX-202, which has altered specificities to both VSCC subtypes. The secondary structures of the three peptides are almost identical, consisting of a triple-stranded beta-sheet and several turns. A comparison of NMR data emphasizes the structural similarities between the peptides and highlights some minor structural differences. In the three-dimensional structures of SVIB and MVIIA these are manifested as orientational differences between two key loops. The structural rigidity of MVIIA was also examined. H alpha shifts are similar in a range of solvents, indicating that there are no solvent-induced changes in structure. The omega-conotoxins form a consensus structure despite differences in sequence and VSCC subtype specificity. This indicates that the omega-conotoxin macrosites for the N/P/Q-subfamily of VSCCs are related, with specificity for receptor targets being conferred by the positions of functional side-chains on the surface of the peptides.
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| 8914922 |
Inhibition of the trehalose-P synthase of mycobacteria by various antibiotics |
10.1006/abbi.1996.0506. |
Arch Biochem Biophys |
Inhibition of the trehalose-P synthase of mycobacteria by various antibiotics
Abstract
- A number of antibiotics were tested as potential inhibitors of the purified trehalose-P synthase of Mycobacterium smegmatis. Of about 30 compounds tested, 4 (cathomycin, circulin, diumycin, and moenomycin) were active against this enzyme. Thus each of these compounds inhibited the formation of trehalose-P by the purified trehalose-P synthase when either UDP-glucose or GDP-glucose was used as the glucosyl donor. However, preincubation of the synthase with heparin, a polyanion activator of the enzyme when UDP-glucose is used as the substrate, prevented the inhibition by these various antibiotics. Fifty percent inhibition by diumycin and moenomycin occurred at a concentration of about 50 microg/ml (Ki of about 1 x 10(-5) M), but 50% inhibition by cathomycin and circulin required substantially higher concentrations (about 50 to 200 microg/ml). The inhibition by cathomycin, diumycin, and moenomycin was of the competitive type, whereas that by circulin was noncompetitive in nature. However, the inhibition was of a complex nature and the data suggest two different binding sites for these inhibitors. Photoaffinity labeling of the synthase with an azido-UDP-32Pglucose probe was effectively blocked by diumycin, moenomycin, or cathomycin indicating that these inhibitors do interact at the substrate binding site. These antibiotics also inhibited the growth of M. smegmatis when added to cells innoculated into trypticase soy broth. The inhibition of growth was concentration-dependent and directly proportional to the size of the bacterial innoculum. These antibiotics, however, did not inhibit protein synthesis nor did they inhibit the incorporation of mannose into lipid-linked saccharides.
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| 8917507 |
Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3. |
10.1073/pnas.93.23.12845 |
Proc. Natl. Acad. Sci. U.S.A. |
Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3.
Abstract
- YY1 is a mammalian zinc-finger transcription factor with unusual structural and functional features. It has been implicated as a positive and a negative regulatory factor that binds to the CCATNTT consensus DNA element located in promoters of many cellular and viral genes. A mammalian cDNA that encodes a YY1-binding protein and possesses sequence homology with the yeast transcriptional factor RPD3 has been identified. A Gal4 DNA binding domain-mammalian RPD3 fusion protein strongly represses transcription from a promoter containing Gal4 binding sites. Association between YY1 and mammalian RPD3 requires a glycine-rich region on YY1. Mutations in this region abolish the interaction with mammalian RPD3 and eliminate transcriptional repression by YY1. These data suggest that YY1 negatively regulates transcription by tethering RPD3 to DNA as a cofactor and that this transcriptional mechanism is highly conserved from yeast to human.
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| 8918811 |
A 1.8 kb alternative transcript from the human epidermal growth factor receptor gene encodes a truncated form of the receptor. |
10.1093/nar/24.20.4050 |
Nucleic Acids Res. |
A 1.8 kb alternative transcript from the human epidermal growth factor receptor gene encodes a truncated form of the receptor.
Abstract
- The epidermal growth factor receptor (EGFR) is encoded by the c-erbB1 proto-oncogene and plays an important role in the control of cell growth and differentiation. To study the potential growth regulatory role of soluble EGF receptors, we have isolated cDNA clones encoding a truncated, secreted form of the human EGFR. The 5' sequence of this cDNA is identical to the EGFR transcript encoding the full-length receptor through exon 10. The unique 3' sequence encodes two additional amino acid residues before encountering an in-frame stop codon, a poly(A) addition site and a poly(A)+ tail. Sequence comparison with genomic DNA sequences demonstrates that this alternative transcript arises by read-through of a splice donor site. As a result, this transcript encodes a portion of the extracellular ligand-binding domain, but lacks the transmembrane domain and the intracellular tyrosine kinase catalytic domain present in the EGFR. Conditioned medium from transfected fibroblast cells contains a 60 kDa protein that is specifically immunoprecipitated by an EGFR monoclonal antibody. These findings demonstrate that alternative processing of the human EGFR transcript produces a secreted product composed of only the extracellular ligand-binding domain.
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| 8920961 |
Analysis of the disulfide linkage pattern in circulin A and B, HIV-inhibitory macrocyclic peptides |
10.1006/bbrc.1996.1708. |
Biochem Biophys Res Commun |
Analysis of the disulfide linkage pattern in circulin A and B, HIV-inhibitory macrocyclic peptides
Abstract
- Circulin A and B are members of a family of macrocyclic peptides, originally isolated from the tropical tree Chassalia parvifolia, that have been shown to display anti-HIV activity. Complete structural elucidation of these highly constrained peptides was difficult due to their cyclic amide backbone and the presence of six disulfide-linked cysteines. In the present study, the disulfide pairing motif of circulin A and circulin B was determined. Since the circulins were resistant to enzymatic proteolysis, cysteine residue pairings were identified by analysis of the complex mixture of cleavage products that resulted from partial acid hydrolysis of the native peptides. Combined utilization of HPLC, fast atom bombardment mass spectrometry and peptide recognition software ("F-MASS" and "F-LINK" programs) were employed to identify the cleavage products. Thus, we were able to unambiguously identify the disulfide linkage pattern in circulin A and circulin B as Cys1-Cys4, Cys2-Cys5 and Cys3-Cys6, where the numbers on the cystine residues refer to their respective order in the peptides.
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| 8925841 |
A member of the arthropod defensin family from edible Mediterranean mussels (Mytilus galloprovincialis) |
10.1111/j.1432-1033.1996.0302h.x. |
Eur J Biochem |
A member of the arthropod defensin family from edible Mediterranean mussels (Mytilus galloprovincialis)
Abstract
- Plasma from the mussel Mytilus galloprovincialis previously immunized by injecting them with bacteria contains several bactericidal proteins. One protein, MGD-1, was purified by reverse-phase HPLC of supernatant from acidified cell-free hemolymph. Its biological activity is directed against both gram-positive and gram-negative bacteria but it is not cytotoxic towards human erythrocytes nor protozoa. As determined by mass spectrometry, the molecular mass of MGD-1 is 4418 Da. Primary-structure analysis revealed 38 amino acids including 8 cysteines and a modified amino acid residue in position 28. Computer searches unambiguously recognized the signature of an arthropod defensin, but the presence of two extra cysteines and of one modified amino acid suggest that it is a previously unknown member of that family.
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| 8939880 |
Characterization of novel cysteine-rich antimicrobial peptides from scorpion blood |
10.1074/jbc.271.47.29537. |
J Biol Chem |
Characterization of novel cysteine-rich antimicrobial peptides from scorpion blood
Abstract
- We have isolated, from the hemolymph of unchallenged scorpions of the species Androctonus australis, three distinct antimicrobial peptides, which we have fully characterized by Edman degradation, electrospray ionization mass spectrometry, and matrix-assisted laser desorption/ionization mass spectrometry. Two are novel molecules: (i) androctonin, a 25-residue peptide with two disulfide bridges, active against both bacteria (Gram-positive and Gram-negative) and fungi and showing marked sequence homology to tachyplesins and polyphemusins from horseshoe crabs; and (ii) buthinin, a 34-residue antibacterial (Gram-positive and Gram-negative) peptide with three disulfide bridges. The third peptide contains 37 residues and three disulfide bridges and clearly belongs to the family of anti-Gram-positive insect defensins. We have synthesized androctonin and explored its activity spectrum and mode of action.
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| 8945785 |
Conformational analysis of a cyclic heptapeptide, pseudostellarin D by molecular dynamics and Monte Carlo simulations |
10.1248/cpb.44.2177. |
Chem Pharm Bull (Tokyo) |
Conformational analysis of a cyclic heptapeptide, pseudostellarin D by molecular dynamics and Monte Carlo simulations
Abstract
- Restrained molecular dynamics calculation in vacuo using AMBER force field implemented in MacroModel/Batchmin showed a major solution conformation in dimethyl sulfoxide-d6 of pseudostellarin D, cyclo(Gly-Tyr-Gly-Pro-Leu-Ile-Leu-). This is a cyclic heptapeptide isolated from Pseudostellaria heterophylla possessing characteristics of a beta-bulge motif with three intramolecular hydrogen bonds, two beta-turns (one type I at Pro4 and Leu5 residues, and one type II at Leu7 and Gly1 residues) and all trans amide bonds. The solution form of pseudostellarin D, which was homologous to that observed in the solid state, was also supported by Monte Carlo simulation study.
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