Delta opioid receptor selectivity induced by conformational constraints in linear enkephalin-related peptides: 1H 400-MHz NMR study and theoretical calculations

Introduction into the structure of the linear hexapeptide DSLET (Tyr-D-Ser-Gly-Phe-Leu-Thr) or DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr) of tert-butyl groups as constraints different from cyclization leads to a large increase in the selectivity for delta opioid binding site in the case of DSTBULET [Tyr-D-Ser-(OtBu)-Gly-Phe-Leu-Thr] (Ki delta = 6.14 nM; Ki mu = 374 nM) and BUBU [Tyr-D-Ser(OtBu)-Gly-Phe-Leu-Thr(OtBu)] (Ki delta = 4.68 nM; Ki mu = 475 nM) or a loss of affinity for DTTBULET [Tyr-D-Thr(OtBu)-Gly-Phe-Leu-Thr] (Ki delta = 866 nM; Ki mu = 4500 nM). This puzzling behavior is studied here by 400-MHz 1H NMR spectroscopy in DMSO-d6 solution and by theoretical calculations. When DSLET and DTLET are compared, the reduction in energetically accessible phi and psi angles induced by the tert-butyl group in the D-Ser2 residue decreases the degree of freedom in the N-terminal part of the peptides. For DSTBULET and BUBU, the rigidification of the backbone evidenced by the appearance of the large NOE's of Phe4 NH-Gly3 alpha and Gly3 NH-alpha and by the loss of the C7 folding around the D-Ser2 residue found in DSLET could explain the drastic loss of affinity for mu opioid receptors. In DTTBULET, a large change in the spatial orientation around the D-Thr2 (OtBu) residue forces the aromatic rings far from each other.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibition of IAA-induced elongation in Avena coleoptile segments by lead: a physiological and an electron microscopic study.

A high resolution growth measuring apparatus was used to demonstrate the inhibition of auxin-induced cell elongation in oat coleoptile segments (Avena sativa L. var Holden) by lead at concentrations ranging from 2 x 10-6 M to 2 x 10-3 M. The inhibition was immediate, having no measurable lag period. Electron micrographs of lead-treated and control segments revealed that in the treated material, lead became localized as electron-dense granules in the cell walls and in vesicles associated with dictyosomes. These granules were found to be lead hydroxide phosphate by electron diffraction techniques. The possible significance of this localization and identification with regard to phosphatase activity is discussed.     

The quantification of formazans in tissue sections by microdensitometry. I. The use of neotetrazolium chloride

Synopsis This article describes the use of a microdensitometer for the measurement of formazan deposits in tissue sections. Some examples are given to illustrate the various applications of this technique in the assessment of glucose-6-phosphate dehydrogenase activity. These are (1) the separate measurement of the red half-formazan intermediate and purple diformazan of neotetrazolium, and the effect of incubation time on their production, (2) the measurement of activities in different regions of the liver lobule, and the selective effect of phenobarbitone, and (3) the measurement of enzyme activity in individual cartilage cells in normal and osteoarthrosis-prone animals. All activities can be expressed in absolute units as nmol hydrogen/mm³/hr, and thus compared with standard biochemical data. The activities obtained all fall within the range of published values for biochemical systems.Paper given at the Royal Microscopical Society's European Histochemistry Meeting at Nottingham in September 1975.     

Schistosoma mansoni: nucleic acid synthesis in immature females from single-sex infections, paired in vitro with intact males and male segments

1. The object of this study was to see whether stimulation of nucleic acid synthesis in immature females by male Schistosoma mansoni is mediated locally by contact, or is propagated systemically in the female. 2. Immature females perfused from single-sex animal infections were paired for one week in vitro with segments of males cut transversely into thirds; others were paired with intact males, or maintained without males; all were then incubated with [3H]-thymidine or tyrosine. 3. Washed females were bisected transversely and isotope uptake counted separately in the anterior and posterior halves. 4. The halves in contact with cut male segments showed significantly higher uptake of [3H]-thymidine than the non-contact halves, indicating increased DNA synthesis and cell division, but non-contact halves had greater uptake of [3H]-tyrosine. 5. Dot-blot hybridization with a female specific single stranded cDNA failed to detect production of the corresponding mRNA in females paired with male segments.