Transglutaminase Catalyzes the Formation of Sodium Dodecyl Sulfate-Insoluble, Alz-50-Reactive Polymers of τ

Abstract: The gene for tryptophan 2,3-dioxygenase (TDO) heretofore was believed to be expressed only in liver. The data presented here demonstrate that RNA encoding TDO is present in rodent brain. Oligonucleotide primers based on the rat liver TDO cDNA sequence were synthesized and used to amplify RNA derived from mouse whole brain and liver and rat brain regions by the RNA-PCR. Reaction products were purified and subjected to DNA sequencing. Identical sequences were obtained when mouse whole brain and liver RNAs were amplified, and these sequences were shown to be 96% identical to the published rat liver tryptophan TDO cDNA sequence. In addition, TDO sequences were found in RNA derived from rat brainstem, cerebellum, cortex, hypothalamus, and the remainder of the brain.     

Sequencing and model structure of a Naja naja atra protein fragment.

We report the amino acid sequence of a basic protein isolated from the snake venom of Naja naja atra. An automated Edman sequencer was used to determine the 65-residue sequence, aided by electrospray ionization/mass spectrometry. Online reduction and pyridylethylation of the peptide was performed to identify the cysteine residues. Trypsin, chymotrypsin and aspartic digestions were carried out to derive peptide fragments for further sequencing. Fragmented peptides were overlapped to obtain the complete sequence. Molecular mass measurements of the whole protein and its fragments were used as a countercheck for sequence assignment. Further confirmation of the sequence was indicated by sequence homology to other snake venom neurotoxins. A molecular model of the tertiary structure was constructed based on sequence homology, and was refined by global minimization and extensive quality control algorithms. Electrostatic and hydrophobic surface calculations and molecular dynamics simulations were carried out to determine the functional properties of the molecule.     

Preferred conformation of the benzyloxycarbonyl-amino group in peptides

Structural parameters, derived from X-ray crystallographic data, have been compiled for 35 derivatives of amino acids, peptides, and related compounds, which contain the N-terminal benzyloxycarbonyl (Z) group. The geometry of the urethane moiety of this end group is closely similar to that of the tert-butoxycarbonyl (Boc) group, except for a relaxation of some bond angles because the Z group is sterically less crowded than the Boc group. For the same reason, the Z group has greater conformational flexibility. As a result, packing forces in the crystal may cause greater deformations of bond angles, resulting in larger variations of observed bond lengths and bond angles than in Boc-peptide crystals. The aromatic rings of the Z end groups tend to stack in crystals. Conformational energy calculations indicate that most conformations of Z-amino acid-N'-methylamides and of corresponding Boc derivatives have similar dihedral angles and relative energies, i.e. the nature of the N-terminal end group has little effect on the conformational preferences of the residue next to it. In particular, the computed fraction of molecules with a cis urethane (C-N) bond is similar for the two derivatives: 0.51 and 0.42 in Boc-Pro-NHCH3 and Z-Pro-NHCH3, respectively, and 0.02 in the two Ala derivatives. There exist several computed conformations of Z-Ala-NHCH3 and Z-Pro-NHCH3 in which the phenyl ring and the C-terminal methylamide group are close to each other. Because of favorable nonbonded interactions, such conformations are of low energy.     

The visual response from the compound eye of Oncopeltus fasciatus: Effects of temperature and sensory adaptation

The effects of temperature (10–30°C) on response latency and amplitude were determined in the dark- and light-adapted compound eye of the milkweed bug, Oncopeltus fasciatus. Response amplitude was an inverse function of temperature under dark-adapted conditions, but was nearly independent of temperature when superimposed on a background field. Response amplitude to a test stimulus (S₂) was maximum at 15–20°C when presented 10 sec after an adapting stimulus (S₁). The optimum temperature (10°C or less) for maximum response amplitude from dark-adapted eyes was far below the temperature at which the animals were grown (19–25°C) and lower than previously reported for insect compound eyes. These results are discussed in terms of the adaptive implications for diurnal, terrestrial ectotherms.Response latency from dark-adapted eyes was an inverse exponential function of temperature with an apparent activation energy of 13·6 kcal mole⁻¹. No change in activation energy could be attributed to light adaptation.Reductions of S₂ response latency and amplitude, during the adaptation régimes employed in this study, were different functions of temperature. Adapting stimuli, which were equally effective at reducing S₂ latency, had different effects on S₂ response amplitude. Recovery of S₂ response latency and amplitude were not related at either 20 or 10°C. Therefore, changes in S₂ response latency as a function of adaptation appeared independent of changes in S₂ response amplitude.