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Information on EC 1.14.13.225 - F-actin monooxygenase and Organism(s) Homo sapiens and UniProt Accession Q8TDZ2
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1.14.13.225 F-actin monooxygenaseIUBMB Comments
The enzyme, characterized from the fruit fly Drosophila melanogaster, is a multi-domain oxidoreductase that acts as an F-actin disassembly factor. The enzyme selectively reduces two L-Met residues of F-actin, causing fragmentation of the filaments and preventing repolymerization . Free methionine is not a substrate . The reaction is stereospecific and generates the (R)-sulfoxide . In the absence of substrate, the enzyme can act as an NAD(P)H oxidase (EC 1.6.3.1) [4,5].
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The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
[F-actin]-L-methionine
+
+
+
=
[F-actin]-L-methionine-(R)-S-oxide
+
+
Synonyms
mical, mical2, mical1, mical-1, mical3, mical-2, mical2pv, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
MICAL1
MICAL
MICAL
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-
-
-
SYSTEMATIC NAME
IUBMB Comments
[F-actin]-L-methionine,NADPH:O2 S-oxidoreductase
The enzyme, characterized from the fruit fly Drosophila melanogaster, is a multi-domain oxidoreductase that acts as an F-actin disassembly factor. The enzyme selectively reduces two L-Met residues of F-actin, causing fragmentation of the filaments and preventing repolymerization [1]. Free methionine is not a substrate [2]. The reaction is stereospecific and generates the (R)-sulfoxide [3]. In the absence of substrate, the enzyme can act as an NAD(P)H oxidase (EC 1.6.3.1) [4,5].
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
[F-actin]-L-methionine + NADH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NAD+ + H2O
-
-
-
?
[F-actin]-L-methionine + NADPH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NADP+ + H2O
-
-
-
?
[F-actin]-L-methionine + NADPH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NADP+ + H2O
-
only 2-3 residues are oxidized in each actin monomer. Several residues, including Met44 and Met47, are modified with similar probability
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?
[F-actin]-L-methionine + NADPH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NADP+ + H2O
-
-
-
?
[F-actin]-L-methionine + NADPH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NADP+ + H2O
usage of F-actin from rabbit skeletal muscle purified from G-actin by ultracentrifugation
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-
?
additional information
?
-
the Km for NADPH of the NADPH oxidase reaction is sensitive to ionic strength and type of ions. The apparent kcat (pH 7) is limited by enzyme reduction by NADPH, which occurs without detectable intermediates. The reaction is also sensitive to solvent viscosity
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-
?
additional information
?
-
-
the Km for NADPH of the NADPH oxidase reaction is sensitive to ionic strength and type of ions. The apparent kcat (pH 7) is limited by enzyme reduction by NADPH, which occurs without detectable intermediates. The reaction is also sensitive to solvent viscosity
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-
?
additional information
?
-
MICAL has additionally NADPH oxidase activity that underlies F-actin disassembly simultaneously with the oxidation of NADPH. For MICAL3 containing the FMO and CH domains, the kcat/Km ratio for NADPH oxidase activity increases dramatically on adding F-actin, while the kcat/Km value for MICAL3 lacking the CH domain changes little
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-
-
additional information
?
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-
MICAL has additionally NADPH oxidase activity that underlies F-actin disassembly simultaneously with the oxidation of NADPH. For MICAL3 containing the FMO and CH domains, the kcat/Km ratio for NADPH oxidase activity increases dramatically on adding F-actin, while the kcat/Km value for MICAL3 lacking the CH domain changes little
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
[F-actin]-L-methionine + NADPH + O2 + H+
[F-actin]-L-methionine-(R)-S-oxide + NADP+ + H2O
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
F-actin
stimulates NADPH oxidation by increasing kcat and kcat/KNADPH about 5 and 200fold, respectively
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additional information
the catalytic efficiency of MICAL3 increases on adding F-actin only when the CH domain is available
-
additional information
-
the catalytic efficiency of MICAL3 increases on adding F-actin only when the CH domain is available
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.134
NADPH
truncated form comprising the MO and CH domains, pH 7.0, 25°C
0.23
NADPH
truncated form comprising the MO, CH and LIM domains, pH 7.0, 25°C
0.5438
[F-actin]-L-methionine
recombinant MICAL3 containing the FMO and CH domains, pH 7.5, 25°C
-
0.8172
[F-actin]-L-methionine
recombinant MICAL3 lacking the CH domain, pH 7.5, 25°C
-
1.438
[F-actin]-L-methionine
pH 8.0, 25°C, recombinant hMICAL3FMOCHDELTA213,530
-
additional information
NADPH
the Km for NADPH of the NADPH oxidase reaction is sensitive to ionic strength and type of ions. pH 7.0, 25°C
additional information
NADPH
-
the Km for NADPH of the NADPH oxidase reaction is sensitive to ionic strength and type of ions. pH 7.0, 25°C
additional information
additional information
Michaelis-Menten kinetics
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.5
NADPH
truncated form comprising the MO, CH and LIM domains, pH 7.0, 25°C
0.0024
[F-actin]-L-methionine
recombinant MICAL3 lacking the CH domain, pH 7.5, 25°C
-
0.0033
[F-actin]-L-methionine
pH 8.0, 25°C, recombinant hMICAL3FMOCHDELTA213,530
-
0.0169
[F-actin]-L-methionine
recombinant MICAL3 containing the FMO and CH domains, pH 7.5, 25°C
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
15.3
NADPH
truncated form comprising the MO, CH and LIM domains, pH 7.0, 25°C
0.0023
[F-actin]-L-methionine
pH 8.0, 25°C, recombinant hMICAL3FMOCHDELTA213,530
-
0.0029
[F-actin]-L-methionine
recombinant MICAL3 lacking the CH domain, pH 7.5, 25°C
-
0.031
[F-actin]-L-methionine
recombinant MICAL3 containing the FMO and CH domains, pH 7.5, 25°C
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
MICAL1 gene disruption in MDA-MB-231 cells knocks out protein expression, which affects F-actin organization, cell size and motility. MICAL1 deletion significantly affects the expression of over 700 genes, with the majority being reduced in their expression levels. Receptor regulator activity is the most significant negatively enriched molecular function gene set. MICAL1 deletion on is also associated with changes in the expression of several serum-response factor regulated genes. MICAL1 disruption attenuates breast cancer tumour growth in vivo
malfunction
the catalytic efficiency of MICAL3 increases on adding F-actin only when the CH domain is available. But this does not occur when two residues, Glu213 and Arg530, are mutated in the FMO and CH domains, respectively
physiological function
additional information
physiological function
MICAL isozymes are involved in actin cytoskeleton reorganization through methionine oxidation. The enzyme functions in F-actin disassembly
physiological function
splicing isoform MICAL2PV is a tunneling nanotubes (TNT) regulator that suppresses TNT formation and modulates mitochondrial distribution. MICAL2PV interacts with mitochondrial Rho GTPase Miro2 and regulates subcellular mitochondrial trafficking. Downregulation of MICAL2PV enhances survival of cells treated with chemotherapeutical drugs. The monooxygenase domain of MICAL2PV is required for its activity to inhibit TNT formation by depolymerizing F-actin
additional information
the catalytic efficiency of MICAL3 increases on adding F-actin only when the CH domain is available. MICAL3 is structurally highly similar to isozyme MICAL1, which suggests that they may adopt the same catalytic mechanism, but the difference in the relative position of the CH domain produces a difference in F-actin substrate specificity. Interaction analysis of the binding site between the CH domain and the FMO domain in human MICAL3, modeling, overview. The FMO-CH interaction in hMICAL3 is required to increase the catalytic efficiency by conferring specific binding to F-actin. The FMO domain that exhibits monooxygenase activity is localized at the N-terminus of MICAL and is highly conserved among species. The CH domain that is usually found in actin binding proteins is adjacent to the FMO domain and is also highly conserved. CH domains are classified into three types: types 1, 2, and 3. Whereas type 3 CH domains are mainly found in regulatory proteins associated with muscle contraction and signaling proteins, type 1 and 2 CH domains are usually found in cytoskeletal proteins. MICALs have a typical type 2 CH domain
additional information
-
the catalytic efficiency of MICAL3 increases on adding F-actin only when the CH domain is available. MICAL3 is structurally highly similar to isozyme MICAL1, which suggests that they may adopt the same catalytic mechanism, but the difference in the relative position of the CH domain produces a difference in F-actin substrate specificity. Interaction analysis of the binding site between the CH domain and the FMO domain in human MICAL3, modeling, overview. The FMO-CH interaction in hMICAL3 is required to increase the catalytic efficiency by conferring specific binding to F-actin. The FMO domain that exhibits monooxygenase activity is localized at the N-terminus of MICAL and is highly conserved among species. The CH domain that is usually found in actin binding proteins is adjacent to the FMO domain and is also highly conserved. CH domains are classified into three types: types 1, 2, and 3. Whereas type 3 CH domains are mainly found in regulatory proteins associated with muscle contraction and signaling proteins, type 1 and 2 CH domains are usually found in cytoskeletal proteins. MICALs have a typical type 2 CH domain
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MICA1_HUMAN
1067
0
117875
Swiss-Prot
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
human MICAL3 contains the flavin-containing monooxygenase (FMO) and calponin-homology (CH) domains, and has an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity like MICAL1, structure comparisons of isozymes MICAL3 and MICAL1. The FMO and CH domains of both isozymes MICAL3 and MICAL1 are highly similar in structure, but superimposition of the two structures shows a different relative position of the CH domain in the asymmetric unit. The catalytic efficiency of MICAL3 dramatically increases on adding F-actin only when the CH domain is available
additional information
-
human MICAL3 contains the flavin-containing monooxygenase (FMO) and calponin-homology (CH) domains, and has an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity like MICAL1, structure comparisons of isozymes MICAL3 and MICAL1. The FMO and CH domains of both isozymes MICAL3 and MICAL1 are highly similar in structure, but superimposition of the two structures shows a different relative position of the CH domain in the asymmetric unit. The catalytic efficiency of MICAL3 dramatically increases on adding F-actin only when the CH domain is available
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
MICAL3 has an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity. The flavin-containing monooxygenase (FMO) and calponin-homology (CH) domains of both MICAL3 and MICAL1 are highly similar in structure, but a different relative position of the calponin-homology domain in the asymmetric unit
purified recombinant His-tagged wild-type and mutant MICAL3 variants, sitting drop vapor diffusion method, method optimization, mixing of 500 nl of 25 mg/ml protein in 50 mM Tris, pH 8.5, 100 mM NaCl, 1 mM 1,4-dithiothreitol, 1% glycerol, with 500 nl of crystallization solution containing 0.1 M bicine-NaOH, pH 9.2, 7% v/v MPD, one day, X-ray diffraction structure determination and analysis at 1.9-2.3 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
expression of truncated forms lacking the C-terminal, LIM and/or calponin homology CH regions. The CH, LIM and C-terminal regions lead to an increase of KmNADPH. The C-terminus also determines an about 10fold decrease of kcat
additional information
construction of enzyme mutants hMCAL3FMOCH, hMICAL3FMO and hMICAL3FMOCHDELTA213,530. The truncated form containing the FMO and CH domains is much more soluble than the full-length form but still retains catalytic activity for F-actin disassembly
additional information
-
construction of enzyme mutants hMCAL3FMOCH, hMICAL3FMO and hMICAL3FMOCHDELTA213,530. The truncated form containing the FMO and CH domains is much more soluble than the full-length form but still retains catalytic activity for F-actin disassembly
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta 2 pLysS by nickel affinity chromatography, ultrafiltration, and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene MICAL3, hMICAL3FMOCH, hMICAL3FMO, and hMICAL3FMODCHELTA213,530 are cloned into the pET-28b plasmid, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 pLysS
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
elevated MICAL1 gene expression is observed in invasive breast cancer samples from human patients relative to normal tissue, while MICAL1 amplification or point mutations are associated with reduced progression free survival
synthesis
-
protocol for obtaining high levels of recombinant protein for the redox only portion of Mical. Cold adapted chaperonins alone do not work well for expressing the Mical redox domain, while the use and removal of solubility tags destabilizes. Low-temperature expression, chaperonins and no solubility tag lead to enhanced expression
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Zucchini, D.; Caprini, G.; Pasterkamp, R.J.; Tedeschi, G.; Vanoni, M.A.
Kinetic and spectroscopic characterization of the putative monooxygenase domain of human MICAL-1
Arch. Biochem. Biophys.
515
1-13
2011
Homo sapiens (Q8TDZ2), Homo sapiens
Vitali, T.; Maffioli, E.; Tedeschi, G.; Vanoni, M.A.
Properties and catalytic activities of MICAL1, the flavoenzyme involved in cytoskeleton dynamics, and modulation by its CH, LIM and C-terminal domains
Arch. Biochem. Biophys.
593
24-37
2016
Homo sapiens (Q8TDZ2)
McGarry, D.J.; Armstrong, G.; Castino, G.; Mason, S.; Clark, W.; Shaw, R.; McGarry, L.; Blyth, K.; Olson, M.F.
MICAL1 regulates actin cytoskeleton organization, directional cell migration and the growth of human breast cancer cells as orthotopic xenograft tumours
Cancer Lett.
519
226-236
2021
Homo sapiens (Q8TDZ2), Homo sapiens
Wang, F.; Chen, X.; Cheng, H.; Song, L.; Liu, J.; Caplan, S.; Zhu, L.; Wu, J.Y.
MICAL2PV suppresses the formation of tunneling nanotubes and modulates mitochondrial trafficking
EMBO Rep.
22
e52006
2021
Homo sapiens (O94851)
Yoon, J.; Wu, H.; Hung, R.J.; Terman, J.R.
Enhanced production of the Mical redox domain for enzymology and F-actin disassembly assays
Int. J. Mol. Sci.
22
1991
2021
Homo sapiens
Kim, J.; Lee, H.; Roh, Y.J.; Kim, H.U.; Shin, D.; Kim, S.; Son, J.; Lee, A.; Kim, M.; Park, J.; Hwang, S.Y.; Kim, K.; Lee, Y.K.; Jung, H.S.; Hwang, K.Y.; Lee, B.C.
Structural and kinetic insights into flavin-containing monooxygenase and calponin-homology domains in human MICAL3
IUCrJ
7
90-99
2020
Homo sapiens (Q7RTP6), Homo sapiens
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