The 28-amino-acid neuropeptide, vasoactive intestinal peptide (VIP), is a potent mitogen during embryonic development and plays a vital role in brain growth. VIP is also mitogenic for tumor cells, including the human neuroblastoma (NMB). Northern blot analysis has revealed VIP mRNA transcripts in NMB. We now report VIP-like immunoreactivity within these neuroblastoma cells that increased during logarithmic growth and decreased after attaining confluency. About 10(6) seeded cells secreted 5-40 pg of VIP-like immunoreactivity into the medium. These results suggest an autocrine role for VIP in the regulation of neuroblastoma growth. A VIP hybrid antagonist (neurotensin6-11 VIP7-28) that has been shown to inhibit lung cancer proliferation was now tested for inhibition of neuroblastoma growth. Receptor binding studies indicated that the hybrid antagonist displaced [125I]-VIP binding in the neuroblastoma cells (EC50 = 5 x 10(-6)M). Furthermore, as measured by thymidine incorporation and by cell counts, the potent VIP hybrid antagonist inhibited neuroblastoma multiplication in a dose-dependent manner. In conclusion, VIP may be an important regulator of growth of nerve cell progenitors and of tumors derived from neuronal origin and intervening with VIP function may lead to improved treatment of cancer.

Female↗

PubMedDate not supplied

Studies toward the biosynthesis of vasoactive intestinal peptide (VIP).

In view of the potential biological importance of VIP, we have begun to examine the regulation of its biosynthesis. For this purpose we have, as a first step, searched for an enriched source of VIP biosynthesis. By a combination of chromatographic procedures and radioimmunoassays we discovered an as yet unknown source for VIP production, namely a human buccal tumor, containing 0.67 +/- 0.05 ng VIP/micrograms protein which is greater than the richest source in brain (the cerebral cortex). Thus, we decided to use the tumor tissue for VIP-mRNA purification and characterization. To identify VIP-mRNA we are using as hybridization probes, synthetic oligodeoxynucleotides with relatively unambiguous nucleotide sequence complementary to the predicted VIP-mRNA sequence. These probes are synthesized, using the deoxynucleoside phosphoramidite approach, to a length of 17 bases each, and contain all the possible DNA sequences according to the genetic code. These specific probes are then radioactively labelled using the reaction catalyzed by the enzyme polynucleotide kinase and afterwards hybridized to mRNA, which had been resolved on denaturing agarose gels. Employing this approach, we identified a single putative VIP-mRNA band which was then partially purified by sucrose gradient centrifugation. Upon in vitro translation in a rabbit reticulocyte lysate cell free system, this mRNA was found to code for VIP immunoreactive proteins. In conclusion, our studies suggest the existence of high molecular weight precursors to VIP cross-reactive with anti-VIP antibodies, that are coded for by a partially purified mRNA containing VIP sequences.

Base Sequence↗

PubMedDate not supplied

Immunoreactive and bioactive somatostatin-like material is present in tobacco (Nicotiana tabacum).

Immunoreactive and biologically active somatostatin-like material is present in extracts of tobacco plants (Nicotiana tabacum). Comparative chromatographic and immunological characterization of this peptide along with synthetic (hypothalamic-like) somatostatin-14 and -28, indicates that both molecular forms are present in the plant. Tobacco-somatostatin inhibits the prostaglandin E2-induced release of growth hormone from cultured anterior pituitary cells. This finding raises questions concerning the evolutionary mechanisms responsible for the presence of this neuropeptide in plants, and the physiological significance of this phenomena.

Animals↗

PubMedDate not supplied

Synthetic peptides from C-reactive protein containing tuftsin-related sequences.

Peptides containing Lys-Pro-Arg or Thr-Lys-Arg segments corresponding to various regions of human C-reactive protein were synthesized. The peptides prepared were composed of amino acid residues, 37-58, 51-58, 173-187 and 181-187 of C-reactive protein. The relationship between C-reactive protein, its synthetic fragments and tuftsin (Thr-Lys-Pro-Arg) was investigated in binding studies, enhancement of phagocytosis and change in cyclic nucleotide levels of mouse macrophages. The peptides AA 51-58 and 181-187 did enhance macrophage phagocytosis capacity to a similar extent to that of tuftsin. They showed however only negligible binding to the cells. The effect of C-reactive protein and the synthetic peptides on metabolic activity of neutrophils was also investigated. It was shown that the peptides inhibited to some degree superoxide production, lysozyme release and Vitamin B12 binding protein release from neutrophils in the absence and presence of the stimulants, PMA or Con A. Comparable activity with tuftsin was not found.

Amino Acid Sequence↗

PubMedDate not supplied

Vasoactive intestinal peptide antagonist retards the development of neonatal behaviors in the rat.

Based on the demonstrated neurotrophic activity of VIP in vitro, a recently designed VIP antagonist was used to assess the role of this neuropeptide in the behavioral development of rats. Rats received daily subcutaneous injections from birth to day 14. Observations of developmental milestones/behaviors were made daily for 21 days. Of the measures of behavioral development tested, the time to surface right on day 4 and the day of onset for forelimb placing, hindlimb placing, forelimb grasping and air righting were significantly retarded by the antagonist. Cotreatment with VIP prevented the antagonist-induced delay. These results suggest that VIP activity is important in the development of select complex motor behaviors.

Animals↗

PubMedDate not supplied

Synthetic peptide approach to the analysis of kinase activities of avian EGF receptor and v-erbB protein.

Analysis of the structure and function of a protein such as the epidermal growth factor receptor is facilitated by the use of antibodies directed against discrete portions of the protein. Here, we describe the characterization and use of antibodies directed against synthetic peptides corresponding to specific portions of the epidermal growth factor receptor and/or v-erbB protein. In particular, one useful antiserum has allowed us to compare the protein kinase activities of the epidermal growth factor receptor and the v-erbB proteins and to conclude that the v-erbB protein is a protein-tyrosine specific kinase as is its homologue the avian epidermal growth factor receptor.

Alpharetrovirus↗