MICAL1 is a cytoplasmic 119 kDa protein participating in cytoskeleton dynamics through the NADPH-dependent oxidase and F-actin depolymerizing activities of its N-terminal flavoprotein domain, which is followed by calponin homology (CH), LIM domains and a C-terminal region with Pro-, Glu-rich and coiled-coil motifs. MICAL1 and truncated forms lacking the C-terminal, LIM and/or CH regions have been produced and characterized. The CH, LIM and C-terminal regions cause an increase of Km,NADPH exhibited by the NADPH oxidase activity of the flavoprotein domain, paralleling changes in the overall protein charge. The C-terminus also determines a ∼10-fold decrease of kcat, revealing its role in establishing an inactive/active conformational equilibrium, which is at the heart of the regulation of MICAL1 in cells. F-actin lowers Km,NADPH (10-50 μM) and increases kcat (10-25 s-1) to similar values for all MICAL forms. The apparent Km,actin of MICAL1 is ∼10-fold higher than that of the other forms (3-5 μM), reflecting the fact that F-actin binds to the flavoprotein domain in the MICAL's active conformation and stabilizes it. Analyses of the reaction in the presence of F-actin indicate that actin depolymerization is mediated by H2O2 produced by the NADPH oxidase reaction, rather than due to direct hydroxylation of actin methionine residues.
Properties and catalytic activities of MICAL1, the flavoenzyme involved in cytoskeleton dynamics, and modulation by its CH, LIM and C-terminal domains / T. Vitali, E. Maffioli, G. Tedeschi, M.A. Vanoni. - In: ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS. - ISSN 0003-9861. - 593(2016 Mar 01), pp. 24-37.
|Titolo:||Properties and catalytic activities of MICAL1, the flavoenzyme involved in cytoskeleton dynamics, and modulation by its CH, LIM and C-terminal domains|
VITALI, TERESA (Primo)
MAFFIOLI, ELISA MARGHERITA (Secondo)
TEDESCHI, GABRIELLA (Penultimo)
VANONI, MARIA ANTONIETTA (Corresponding)
|Parole Chiave:||Cytoskeleton; F-actin depolymerization; FAD-containing monooxygenase/oxidase; Flavoprotein; MICAL1; Semaphorin signaling; Biochemistry; Biophysics; Molecular Biology|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
|Data di pubblicazione:||1-mar-2016|
|Data ahead of print / Data di stampa:||1-feb-2016|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.abb.2016.01.016|
|Appare nelle tipologie:||01 - Articolo su periodico|