• Characterization of anti-myosin monoclonal antibodies.

      Nelson, Paul N.; Astley, S.J.; Roden, Denise A.; Waldron, E.E.; Baig, K.M.; Caforio, A.L.; Koutedakis, Yiannis; Perera, Shantha; Spry, C. (New Rochelle (NY): Mary Ann Liebert, Inc., 2005)
      The characterization of monoclonal antibodies (MAbs) with regard to reactivity and specificity is important for the successful application as a molecular probe and/or diagnostic reagent. Furthermore, it is recognized that some monoclonal reagents perform well in some assay systems but not others. In this study, the reactivity profiles of two anti-myosin MAbs (H1 and DH2, raised against human cardiac myosin) were evaluated in enzyme-linked immunosorbent assay (ELISA), slot-blotting, and immunocytochemistry. Both antibodies performed well in slot-blotting against myosin heavy chain preparations from cardiac and skeletal muscle and from non-human sources. In general, MAb H1 demonstrated strong to moderate reactivity in all assay systems, whilst MAb DH2 faired poorly in ELISA. MAb H1 also showed reactivity to synthetic peptides of myosin, one of which possessed a motif (ERRDA, single amino acid code) that was found in other human and nonhuman myosin protein sequences that could explain its cross-reactive profile. Intriguingly, this motif was found on viral and other pathogenic agents associated with myocarditis. Hence, it is speculated that this region could give some credence to the mechanism of molecular mimicry associated with some cardiac diseases. Overall, MAb H1 may serve as a useful probe of myosin structure.
    • Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel

      Kitchen, P; Day, RE; Taylor, LHJ; Salman, MM; Bill, RM; Conner, Matthew T.; Conner, Alex C.; From the Molecular Organisation and Assembly in Cells Doctoral Training Centre, University of Warwick, Coventry CV4 7AL. (American Society for Biochemistry & Molecular Biology (ASBMB), 2015-05-26)
      © 2015 by The American Society for Biochemistry and Molecular Biology, Inc. Background: The water channel protein aquaporin 4 (AQP4) controls water permeability of the blood-brain barrier. Results: Hypotonicity induces rapid relocalization of AQP4 in a calcium-, calmodulin-, and kinase-dependent manner. Conclusion: AQP4 can be relocalized between the cell membrane and intracellular compartments. Significance: Pharmacological modulation of AQP4 membrane localization could provide a new approach to treating brain edema.