Structural States and Dynamics of the D-Loop in Actin

Conformational changes induced by ATP hydrolysis on actin are involved in the regulation of complex actin networks. Previous structural and biochemical data implicate the DNase I binding loop (D-loop) of actin in such nucleotide-dependent changes. Here, we investigated the structural and conformational states of the D-loop (in solution) using cysteine scanning mutagenesis and site-directed labeling. The reactivity of D-loop cysteine mutants toward acrylodan and the mobility of spin labels on these mutants do not show patterns of an α-helical structure in monomeric and filamentous actin, irrespective of the bound nucleotide. Upon transition from monomeric to filamentous actin, acrylodan emission spectra and electron paramagnetic resonance line shapes of labeled mutants are blue-shifted and more immobilized, respectively, with the central residues (residues 43–47) showing the most drastic changes. Moreover, complex electron paramagnetic resonance line shapes of spin-labeled mutants suggest several conformational states of the D-loop. Together with a new (to our knowledge) actin crystal structure that reveals the D-loop in a unique hairpin conformation, our data suggest that the D-loop equilibrates in F-actin among different conformational states irrespective of the nucleotide state of actin.     

Characterization of aPY-like peptides in anglerfish brain using a novel radioimmunoassay for aPY-Gly

Anglerfish peptide YG (aPY) was isolated from pancreatic islets of the anglerfish. Subsequent immunohistochemical and biochemical analyses demonstrated that anglerfish islet cells synthesize aPY. We have now developed and characterized a radioimmunoassay (RIA) for aPY and have examined extracts of anglerfish brain for aPY-like peptides. Brain extracts were subjected to gel filtration and high performance liquid chromatography (HPLC). Fractions from HPLC eluates were analyzed in the aPY RIA and also in a neuropeptide Y (NPY) RIA. A single peak of aPY-like immunoreactivity eluted from HPLC columns. The elution position of this aPY-like peptide coincided exactly with the aPY-Gly marker under several gradient conditions. Results from the NPY RIA confirmed the presence of several molecular forms of NPY-like immunoreactive peptides in the anglerfish brain. These results demonstrate the utility of the newly developed aPY RIA for studies of anglerfish brain peptides and extend our previous immunohistochemical demonstration of aPY-like staining in the anglerfish brain.     

The importance of oedema in acute radiation injury to the cerebral cortex of rats: An electron microscope study

Summary The oedematous changes in the cerebral cortex of rats after acute irradiation are described. The importance of these changes in the over-all injury is discussed. Swelling and rupture of clear glial cells without expansion of the extracellular space are seen during the early phases of injury. Not all of the clear glia become swollen, but those that do eventually undergo complete dissolution, initiating small foci of severe damage. The necrosis that ultimately encompasses the entire irradiated field may spread from these initial sites of injury. Clear glial cells that do not undergo swelling show an unusual type of reaction before they become necrotic.This work was supported by the U.S. Atomic Energy Commission and formed a part of a dissertation submitted in partial fulfillment for the degree of Doctor of Philosophy to the faculty of the Graduate School of Loyola University, Chicago, Illinois.The author wishes to express his indebtedness to Dr. F. Wassermann of Argonne National Laboratory, and Dr. D. S. Jones of Stritch School of Medicine, Loyola University, for their advice and guidance during the course of this study.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his 80th birthday.