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Biomedical subjects

R W Wallace

Publications and source records attributed to R W Wallace.

51 records · Page 3Linked to original sources
PubMed1980

High levels of a heat-labile calmodulin-binding protein (CaM-BP80) in bovine neostriatum.

Bovine brain contains a heat-labile, 80,000-dalton calmodulin-binding protein (CaM-BP80) which inhibits the calmodulin-dependent activities of cyclic 3',5'-nucleotide phosphodiesterase, adenylate cyclase, and Ca2+-ATPase in vitro. CaM-BP80 is composed of two polypeptides (60,000 and 18,500 daltons) present in a 1:1 ratio. An antibody directed against CaM-BP80 was raised in rabbits, and a radioimmunoassay was developed, having a sensitivity of 60 fmol of CaM-BP80. Using the radioimmunoassay, we determined the levels of CaM-BP80 in various bovine tissues. The protein was found primarily in the brain, present in particularly high levels in the neostriatum. These results, together with immunohistochemical localization of CaM-BP80 at the postsynaptic densities and the microtubules of postsynaptic dendrites [Wood, J.G., Wallace, R., Whitaker, J., & Cheung, W.Y. (1980) J. Cell Biol. 84, 66-76], suggest that the protein may have a role in the cerebrum at the site of neurotransmitter action and at the level of microtubular function.

Animals↗
PubMed1980

Localization of calmodulin in rat tissues.

The localization of calmodulin, a calcium-dependent modulator of many enzymes, was studied in rat liver, skeletal muscle, and adrenal slices. Calmodulin is found in liver cytoplasm, nucleus, and plasma membrane. Much of the cytoplasmic calmodulin is associated with glycogen particles presumably bound to enzymes involved in glycogen metabolism. Skeletal muscle calmodulin is found on the I-band, also associated with glycogen particles. Intermyofibrillar staining that is not glycogen associated is also observed. Calmodulin is localized in the cytoplasm and nucleus of adrenal cortex cells. Injection of corticotropin leads to a greatly increased localization of calmodulin in nuclei of the adrenal cortex. These observations suggest that one role of calmodulin may be the regulation of hormone effects on nuclear processes.

Animals↗
PubMed1980

Immunocytochemical localization of calmodulin and a heat-labile calmodulin-binding protein (CaM-BP80) in basal ganglia of mouse brain.

Antisera to calmodulin, a Ca2%-dependent modulator protein, and a heat-labile calmodulin-binding protein have been used to localize these proteins in mouse caudate-putamen. The two proteins appear to be located at identical sites in this brain area. At the light microscopic level, calmodulin and calmodulin-binding protein are found within the cytoplasm and processes of large cells. At the electron microscopic level the proteins are associated with neuronal elements only, primarily at postsynaptic sites within neuronal somata and dendrites. Within the dendrites the immunocytochemical label is associated predominantly with the postsynaptic density and dendritic microtubules. These results are in accord with recent biochemical and immunihistochemical studies of calmodulin in brain and in dividing cells. Thus, calmodulin and the heat-labile calmodulin-binding protein may play a role in the nervous system at the site of neurotransmitter action and at the level of microtubular function.

Animals↗
PubMed1979

Calmodulin. Production of an antibody in rabbit and development of a radioimmunoassay.

Calmodulin, a heat-stable Ca2+-binding protein (Mr = 16,700) found in all eukaryotes, is a multifunctional modulator, mediating many of the effects of Ca2+ in cellular functions. The protein was derivatized with 1-fluoro-2,4-dinitrobenzene (DNB) to give 3 mol of DNB/mol of calmodulin (DNB3-calmodulin). The dinitrophenylated protein was almost as active as native calmodulin in stimulating bovine brain Ca2+-dependent phosphodiesterase. Incorporation of the dinitrophenyl groups renders calmodulin highly antigenic in the rabbit; native calmodulin is a weak antigen. Rabbits immunized with DNB3-calmodulin produced specific antibody against both DNB3-calmodulin and calmodulin. Using the immunized serum, a radioimmunoassay was developed for calmodulin, the sensitivity for DNB3-calmodulin and calmodulin being approximately 0.2 and 2 pmol, respectively. Although the sensitivity of the radioimmunoassay for calmodulin is comparable to the enzyme assay of calmodulin with Ca2+-dependent phosphodiesterase, the radioimmunoassay affords the detection of calmodulin on the basis of antigenic determinants, and thus measures calmodulin in terms of polypeptide structure instead of its ability to stimulate an enzyme. Further, the accuracy of the radioimmunoassay is not affected by the presence of a heat-labile inhibitor protein, which affects the enzyme assay to give an apparent underestimation.

Animals↗
PubMed1979

Purification and characterization of an inhibitor protein of brain adenylate cyclase and cyclic nucleotide phosphodiesterase.

A heat-labile inhibitor protein of adenylate cyclase (EC 4.6.1.1) and phosphodiesterase (EC 3.1.4.17) has been purified to apparent homogeneity from bovine brain cerebrum by a simple two-column procedure. The inhibitor exerts its effect on adenylate cyclase or phosphodiesterase by forming a complex with the Ca2+-dependent activator protein, thereby competing with the apoenzyme for the activator. The protein was estimated to have a molecular weight of 80,000 and a Stokes radius of 39 A by gel filtration. The inhibitor was resolved in a sodium dodecyl sulfate-polyacrylamide gel into two equal molar subunits, with molecular weights of 60,000 and 18,500. In the presence of the activator and Ca2+, the thermal stability of the inhibitor was increased, indicative of a new conformation. The effectiveness of the inhibitor varied considerably, depending on its sequence of addition to the reaction mixture relative to phosphodiesterase and the activator protein, presumably because the activator appeared to have a greater affinity for the inhibitor than for phosphodiesterase.

Adenylyl Cyclase Inhibitors↗
PubMed1978

Cosmic radiation exposure in subsonic air transport.

This FAA- and NASA-sponsored study of cosmic radiation doses recieved by United States residents flying in commercial jet aircraft is the most extensive to date and combines computer calculations with experimental data. Data derived from 1973 statistics on 2.99 million intercity flights carrying 468 million seats were included in the calculations, yielding a total of 581 billion seat-kilometer. The average flight was 1,084 km in length, was flown at an altitude of 9.47 km, and lasted 1.41 h. The average dose rate was 0.20 mrem/h, resulting in an average passenger dose of 2.82 mrem/year and an average crewmember dose of 160 mrem/year. The average radiation dose to the total U.S. population was 0.47 mrem/person/year. These results are in good agreement with data from several experiments performed by us and others in aircraft at various altitudes and latitudes. The significance of these doses to the population is discussed.

Adolescent↗
PubMedDate not supplied

Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells.

Chloride impermeability of epithelial cells can account for many of the experimental and clinical manifestations of cystic fibrosis (CF). Activation of apical-membrane Cl- channels by cyclic AMP-mediated stimuli is defective in CF airway epithelial cells, despite normal agonist-induced increases in cellular cAMP levels. This defect in Cl- channel regulation has been localized to the apical membrane by exposing the cytoplasmic surface of excised membrane patches to the catalytic subunit (C subunit) of cAMP-dependent protein kinase and ATP. In membranes from normal cells, C-subunit activated Cl- channels with properties identical to those stimulated by cAMP-dependent agonists during cell-attached recording. Activation by the C subunit was not observed in CF membranes, but the presence of Cl- channels was verified by voltage-induced activation. The failure of the C subunit to activate the Cl- channels of CF membranes indicates that the block in their cAMP-mediated activation lies distal to induction of cAMP-dependent protein kinase activity and focuses our attention on the Cl- channel and its membrane-associated regulatory proteins as the probable site of the CF defect.

Cell Membrane↗