Enzymatic and Actin-Binding Properties of MICAL-1

actin;axon guidance;actin depolymerization;cytoskeleton reorganization;enzyme kinetics;flavin-containing monoxygenases;hydrogen peroxide;MICAL;redox signaling;signal transduction;Biophysics

Simon, Olivier BauerAmzel, L. Mario ;

Johns Hopkins University

关键词: actin;    axon guidance;    actin depolymerization;    cytoskeleton reorganization;    enzyme kinetics;    flavin-containing monoxygenases;    hydrogen peroxide;    MICAL;    redox signaling;    signal transduction;    Biophysics;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/40181/SIMON-DISSERTATION-2014.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

MICAL is a large multi-domain monooxygenase with orthologues in flies and humans. MICAL is known to play an important role in axon guidance through semaphorin signaling and, in particular, it has recently been found to influence actin polymerization, with certain constructs capable of disassembling F-actin through methionine oxidation. MICAL also can produce hydrogen peroxide, a potential signaling molecule. In the following, I expressed and purified several proteins, including the MICAL-1 C-terminus, the cytoplasmic portion of the semaphorin receptor Plexin-A1, and the small GTP-binding protein Rnd1, combined these with MICAL constructs of varying length, and used kinetics, co-sedimentation, peroxide quantitation, HPLC, and size exclusion in an effort to characterize the factors necessary to activate MICAL;;s peroxide-producing and actin-disassembly functions.