Peptide and glycopeptide dendrimers. Part I

Recent progress in peptide and glycopeptide chemistry make the preparation of peptide and glycopeptide dendrimers of acceptable purity, with designed structural and immunochemical properties reliable. New methodologies using unprotected peptide building blocks have been developed to further increase possibilities of their design and improve their preparation and separation. Sophisticated design of peptide and glycopeptide dendrimers has led to their use as antigens and immunogens, for serodiagnosis and other biochemical uses including drug delivery. Dendrimers bearing peptide with predetermined secondary structures are useful tools in protein de novo design. This article covers synthesis and applications of multiple antigen peptides (MAPs), multiple antigen glycopeptides (MAGs), multiple antigen peptides based on sequential oligopeptide carriers (MAP‐SOCs), glycodendrimers and template‐assembled synthetic proteins (TASPs). Part I deals with the development of various structural forms of MAPs as well as their application as antigens, immunogens, and for immunodiagnostic and biochemical purposes. Copyright © 1999 European Peptide Society and John Wiley & Sons, Ltd.     

蜜蜂褪黑素的测定方法

在脊椎动物中,褪黑素(5-甲氧基-N-乙酰色胺,MLT)是一种由松果体分泌的具有典型光周期信号作用的神经内分泌激素。近年在昆虫头部也发现了这种化学物质。该文简述了褪黑素常见测定方法,详细介绍了褪黑素的免疫测定方法RIA(radionimmunoassay),并使用该方法初步测定了中华蜜蜂ApisceranaFabricius工蜂头部褪黑素的含量,结果显示褪黑素含量与工蜂的社会分工相关。     

Absence of inactivation or phosphorylation of ornithine decarboxylase by nuclear protein kinase NII and of immunological cross-reactivity between RNA polymerase I and ornithine decarboxylase

Incubation with protein kinase NII did not result in phosphorylation or inactivation of mouse kidney ornithine decarboxylase. Partially purified ornithine decarboxylase preparations contained a protein kinase activity and stimulated the activity of RNA polymerase I. However, these properties were due to contaminating protein(s) since further purification reduced the kinase activity and removal of the ornithine decarboxylase with a specific antiserum did not abolish the ability to stimulate RNA polymerase I. Antibodies to RNA polymerase I did not interact with ornithine decarboxylase and antibodies to ornithine decarboxylase did not interact with RNA polymerase I. These results indicate that: a) mammalian ornithine decarboxylase activity is not regulated by phosphorylation by protein kinase NII or the contaminating kinase, and b) the ability of impure preparations of ornithine decarboxylase to stimulate RNA polymerase I is due to a contaminating unrelated protein.     

虎纹蛙促性腺激素含量随年龄及季节的变化(英文)

促性腺激素（ＧｔＨ;或ＬＨ和ＦＳＨ）在脊椎动物生殖调节中起中Ｃ作用;脑垂体和血浆ＧｔＨ水平在一定程度上反映着动物体的生殖生理状态。在蛙类,对脑垂体和血浆的ＬＨ及ＦＳＨ含量进行过较全面研究的,仅在牛蛙和日本蟾蜍有过报道,结果显示它们的ＬＨ和ＦＳＨ的含量存在有种类差异性。虎纹蛙属中国二级保护动物,也是唯一受保护的蛙类,对其生殖生物学的基础内容进行研究具有重要的意义。 为此,本文利用放射免疫测定法,测定了虎纹蛙幼蛙和不同性腺发育阶段（季节）成蛙脑垂体与血浆的ＬＨ及ＦＳＨ含量,以弄清这些激素的含量变化与年龄、季节（性腺发育阶段）变化的关系,以期为虎纹蛙的基础生殖生物学及蛙类的生殖内分泌学充实新的内容,为虎纹蛙的人工繁殖和保护提供理论依据。结果是：幼蛙血浆ＬＨ水平显著高于各期成蛙（Ｆｉｇ．Ａ）,血浆ＦＳＨ水平显著低于成熟前期成蛙,而和其它各性腺发育阶段成蛙相当（Ｆｉｇ．Ｂ）。而脑垂体ＬＨ或ＦＳＨ的含量显著低于各期成蛙（Ｆｉｇ．Ｃ＆Ｄ）。这说明,幼蛙脑垂体已具有一定的合成和释放ＬＨ及ＦＳＨ的能力。 成蛙脑垂体和血浆ＬＨ及ＦＳＨ水平随性腺发育阶段（季节）的不同而有一定的变化：雌蛙血浆ＬＨ水平在成熟期最高,性腺再发育期最低;     

Expression of hepatitis B virus surface antigen P31 gene in Escherichia coli

A hepatitis B virus surface antigen (HBsAg) P31-coding DNA was constructed from a DNA fragment of the plasmid pHBr330 containing the entire hepatitis B virus (HBV) adr DNA and a chemically synthesized adaptor. The P31 gene was inserted into an expression vector, pTRP771, having an Escherichia coli tryptophan operon (trp) promoter to give a recombinant plasmid pTRP P31-R. The distance between the Shine-Dalgarno (SD) sequence and the initiation codon of P31 gene was adjusted to 9 bp. The expression level of HBsAg by E. coli 294[pTRP P31-R] was significantly elevated, in contrast to that of HBsAg by E. coli 294[pTRP SS-6]. Western blotting analysis has shown that E. coli[pTRP P31-R] synthesizes a specific polypeptide P31 of about 31 kDal, which reacts with anti-HBsAg antibody. The binding studies with polyalbumins from various species have also suggested that HBsAg P31 specifically binds to polymerized human serum albumin.     

Distribution of enteric neural peptide YY in the dog gastrointestinal tract

Various regions of the dog gastrointestinal tract were investigated for the distribution of peptide YY (PYY) neurons using immunocytochemistry and radioimmunoassay. PYY neurons that encircled non-PYY-immunoreactive neurons were mainly observed in the myenteric plexus from the stomach to the colon. There was more PYY-like immunoreactivity in the muscle layer of the stomach and ileum than in the other intestines. The results of high performance liquid chromatography revealed that neural PYY-immunoreactive substance is identical to authentic PYY. PYY was not localized in the cholinergic neurons. These results indicate that PYY, as a neuropeptide, is involved in the regulation of gastrointestinal function.     

Ecdysteroids in nemerteans: further characterization and identification

Ecdysteroids are a class of steroidal hormones thatare important in molting and reproduction inarthropods. These hormones have been recentlydetected in non-arthropodan groups, such assoft-bodied worms. To continue our efforts todetermine the presence of ecdysteroids in nemerteans,this study further documents the identification of20-hydroxyecdysone (20E), in Paranemertesperegrina, by gas chromatography/mass spectrometry(GC/MS). After C₁₈ Sep-Pak extraction,radioimmunoassay of the 40% and 80% methanolicfractions from an extract of 1000 worms indicated 98and 156 ng of ecdysteroids, respectively. Ecdysteroids of these two samples, as well as the 20Eand ecdysone standards, were N-trimethylsilyimidazole(TMSI)-derivatized before GC/MS analysis. Themethanolic samples contained a large number ofcompounds but only small, insignificant peaks in thearea where ecdysteroid standards eluted. However, thereconstruction ion current (RIC) chromatograms for m/z561 indicated the presence of 20E at the correctretention time of 12.48 min in the 40% methanolicfraction. Reanalysis of the samples under selectedion monitoring mode demonstrated the presence of 20Ein both the 40% and 80% fractions. These resultsdemonstrate conclusively the presence of an activeecdysteroid in the phylum Nemertea.     

pH-induced conformational changes in chromatin subunits measured by circular dichroism and immunochemical reactivity

Changes in the conformational state of chromatin core particles from chicken erythrocytes were studied by both immunochemical and biophysical methods as a function of pH and ionic strength. When the pH of core particles in a solution of ionic strength 3, 60 or 220 mM was lowered from pH 7.5, a sharp transition in the circular dichroism spectrum of DNA monitored between 320 and 260 nm was observed at pH 6.65. This change in DNA ellipticity was totally reversible. Binding to core particles of antibodies specific for histones H2B, H2A, H3 and for the IRGERA (synthetic C-terminal) peptide of H3 was used to follow changes in histone antigenicity. Binding was studied in the pH range 7.5-5.35, and at ionic strength of 60 and 220 mM. A change in reactivity of some histone epitopes was observed around pH 6.2–6.5. However, the changes observed by circular dichroism and antibody binding pertain to different components of chromatin subunits and they probably reflect independent phenomena. The alteration in accessibility of these determinants at the surface of core particles was completely reversible and was dependent on ionic strength. The conformation changes in core particles occurring near physiological ionic strength and pH may reflect dynamic changes in chromatin structure that possess functional significance.     

Is caerulein amphibian CCK?

The amphibian skin decapeptide caerulein is structurally related to the mammalian peptides gastrin and CCK, suggesting that the peptides might share a common evolutionary history. It has been suggested that caerulein is the amphibian counterpart of gastrin and CCK, and that the Amphibia do not possess authentic gastric and CCK. High Performance Liquid Chromatography (HPLC) in conjunction with radioimmunoassay using a caerulein-specific antiserum and C-terminal CCK antisera, was used to characterize CCK-and caerulein-like peptides in amphibian brain and gut. In the brain of Xenopus laevis, two CCK-like peptides were present, one of which was indistinguishable by HPLC from mammalian CCK8. No decapeptide caerulein was detected in the brain of Xenopus laevis or Rana temporaria. In the stomach of Xenopus and in the intestine of both species studied, CCK-like and caerulein-like peptides were present. The results indicate therefore that the Amphibia possess CCK8-like rather than caerulein-like peptides in brain. In contrast, stomach and intestine contain both CCK-like and caerulein-like peptides, but the latter are however distinguishable from the decapeptide found in skin.     

Effects of hibernation on somatostatin-like immunoreactivity in the brain of the ground squirrel (Spermophilus richardsonii) and European hedgehog (Erinaceus europaeus)

Summary The localization of the somatostatin system in the brains of Richardson's ground squirrels (Spermophilus richardsonii) and European hedgehogs (Erinaceus europaeus) was described by use of immunocytochemical methods. In addition, (i) chemically differing types of somatostatin and (ii) different activity phases of the somatostatin system during the hibernation cycle were investigated in the ground squirrel by means of high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA).In both species, the hypothalamic component of the somatostatin system (periventricular nuclei, fiber projections to the median eminence) is more prominent than the widespread extrahypothalamic representation of the system displaying mainly scattered perikarya and nerve fibers. The reactivity pattern of the somatostatin system varied among hibernating, aroused, and non-hibernating animals; moreover, the interspecific differences were pronounced. The activity of the hypothalamic somatostatin system in the hibernating ground squirrel appeared to be suppressed when compared to non-hibernating controls, whereas in the hibernating hedgehog this system showed signs of increased activity in comparison to non-hibernating controls. In contrast, in the present material the extrahypothalamic components of the somatostatin system did not exhibit significant changes in their activity.