Information on EC 3.1.3.53 - [myosin-light-chain] phosphatase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY hide
3.1.3.53
-
RECOMMENDED NAME
GeneOntology No.
[myosin-light-chain] phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
[myosin light-chain] phosphate + H2O = [myosin light-chain] + phosphate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
additional information
SYSTEMATIC NAME
IUBMB Comments
[myosin-light-chain]-phosphate phosphohydrolase
The enzyme is composed of three subunits. The holoenzyme dephosphorylates myosin light chains and EC 2.7.11.18, myosin-light-chain kinase, but not myosin; the catalytic subunit acts on all three substrates.
CAS REGISTRY NUMBER
COMMENTARY hide
108658-39-5
-
86417-96-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Madin Darby
-
-
Manually annotated by BRENDA team
Canis lupus familiaris Madin Darby
Madin Darby
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
MLCP
-
-
Manually annotated by BRENDA team
PP-1
-
-
Manually annotated by BRENDA team
PP-1M
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
MLCP
-
-
Manually annotated by BRENDA team
Mus musculus BALB/c
BALB/c
-
-
Manually annotated by BRENDA team
normal and pulmonary hypertensive fetal sheep, MLCP, protein phosphatase type 1
-
-
Manually annotated by BRENDA team
Rattus norvegicus Sprague-Dawley
Sprague-Dawley
-
-
Manually annotated by BRENDA team
Sus scrofa SMPP-1M
SMPP-1M
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
phosphorylation of MYPT1 is a major mechanism of MLCP regulation, but protein-protein interactions may also be important. Ca2+-dependent and Rho-associated kinase-mediated regulation of myosin light chain kinase and myosin light chain phosphatase, respectively, in the arterial myogenic response, molecular mechanisms, overview
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
histone deacetylase 7 phosphate + H2O
histone deacetylase 7 + phosphate
show the reaction diagram
myosin light-chain phosphate + H2O
myosin light-chain + phosphate
show the reaction diagram
myosin regulatory light-chain phosphate + H2O
myosin regulatory light-chain + phosphate
show the reaction diagram
P-MLC20 + H2O
MLC20 + phosphate
show the reaction diagram
-
P-MLC20: phospho-peptide mimicking MLC20(3-26:P-Ser19)
-
-
?
phosphorylated glycogen synthase + H2O
glycogen synthase + phosphate
show the reaction diagram
phosphorylated heavy meromyosin + H2O
heavy meromyosin + phosphate
show the reaction diagram
phosphorylated myosin + H2O
myosin + phosphate
show the reaction diagram
phosphorylated myosin-light chain kinase + H2O
myosin light-chain kinase + phosphate
show the reaction diagram
phosphorylated phoshorylase A + H2O
phosphorylase A + phosphate
show the reaction diagram
phosphorylated phoshorylase B + H2O
phosphorylase B + phosphate
show the reaction diagram
-
myosin-associated PP-1M
-
?
phosphorylated phosphorylase kinase + H2O
phosphorylase kinase + phosphate
show the reaction diagram
radixin + H2O
?
show the reaction diagram
regulatory myosin light-chain phosphate + H2O
regulatory myosin light-chain + phosphate
show the reaction diagram
[myosin light-chain] phosphate + H2O
[myosin light-chain] + phosphate
show the reaction diagram
[myosin regulatory light chain] phosphate + H2O
[myosin regulatory light chain] + phosphate
show the reaction diagram
-
from chicken gizzard
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
myosin light-chain phosphate + H2O
myosin light-chain + phosphate
show the reaction diagram
phosphorylated myosin + H2O
myosin + phosphate
show the reaction diagram
phosphorylated myosin-light chain kinase + H2O
myosin light-chain kinase + phosphate
show the reaction diagram
-
SMP-I modulates the activity of myosin-light chain kinase
-
?
radixin + H2O
?
show the reaction diagram
-
an ERM protein
-
-
?
[myosin light-chain] phosphate + H2O
[myosin light-chain] + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
-
highly activating at 0.0016 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(S)-2-methyl-1-[(4-methyl-5-isoquinolinyl)sulfonyl]-hexahydro-1H-1,4-diazepine
-
H-1152, 0.0003 mM produces a strong inhibition of basal MYPT1-pT853 by nearly 75%, KCl produces an approximate 1.75-2fold increase in MYPT1-pT853 in the presence and absence of H-1152
14-3-3beta protein
-
70% inhibition in the presence of 0.5 mM 14-3-3beta, dissociates MLCP from myosin II
-
acidic phospholipid
-
interaction of enzyme with acidic phospholipids inhibits activity toward phosphorylated myosin
-
ADP
-
SMP-I, catalytic subunit is more sensitive than holoenzyme
AMP
-
SMP-I, catalytic subunit is more sensitive than holoenzyme
arachidonic acid
-
inhibits SMPP-1M, dissociates the catalytic subunit from the native holoenzyme, inhibits the regulatory action of the 110 kDa/21 kDa subunit complex on the catalytic subunit PP1C activity, the C-terminal half of the 110 kDa subunit is required for inhibition
calyculin
CPI-17
dephosphorylated myosin light chain
-
SMP-I, product inhibition
diphosphate
flavone
-
inhibits U46619-induced regulatory myosin light-chain phosphorylation
fragment 654-880 of human myosin phosphatase targeting subunit GST-fusion
-
-
-
fragment 697-880 of human myosin phosphatase targeting subunit GST-fusion
-
-
-
glycerol
-
inhibits SMMP activity, dissociates P-myosin primarily into an extended conformation
KCl
-
at high concentrations
MgATP2-
-
inhibits SMMP activity towards P-myosin, disassembles P-myosin mostly into a folded conformation
microcystin
-
-
-
Mn2+
-
at 1 mM inhibitory effect
okadaic acid
phosphate
-
SMP-I, holoenzyme and catalytic subunit, product inhibition
phosphatidic acid
-
most effective inhibitor among acidic phospholipids, followed by phosphatidylserine and phosphatidylinositol, phosphorylated myosin as substrate
phosphatidylinositol
-
phosphatidic acid is the most effective inhibitor among acidic phospholipids, followed by phosphatidylserine and phosphatidylinositol, phosphorylated myosin as substrate
phosphatidylserine
-
phosphatidic acid is the most effective inhibitor among acidic phospholipids, followed by phosphatidylserine and phosphatidylinositol, phosphorylated myosin as substrate
phospho-CPI-17
Rho/Rho kinase
smoothelin-like 1 protein
-
originally called CHASM (calponin homology-associated smooth muscle), 55% inhibition at 0.0025 mM, 75% inhibition at up to 0.02 mM
-
tautomycetin
-
type 1 phosphatase inhibitor
tautomycin
-
PP-1M inhibitor
thrombin
-
activates Rho/Rho kinase to inactivate PP-1M
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KCl
-
in organ culture
p116Rip
-
RhoA-binding protein, activates enzyme activity of the holoenzyme due to binding to MYPT 1 targeting subunit. Activation is specific to myosin as substrate
-
sodium nitroprusside
-
nitrovasodilator, increases MLCP activity in intact carotid media, mechanism
type I cGMP-dependent protein kinase
-
-
-
U46619
-
thromboxane A2 analogue at 10-7 mol/L, increases phosphorylation of MYPT1 and regulatory myosin light-chain
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0059
heavy meromyosin
-
pH 7, 30C, enzyme SMP-IV
-
0.0015 - 0.05
myosin light-chain
0.00103 - 0.01117
myosin light-chain phosphate
0.00045 - 0.0024
Phosphorylated myosin
0.01
[myosin light-chain] phosphate
-
pH 7.6, temperature not specified in the publication
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0115
dephosphorylated myosin light chain
-
pH 7, 30C, SMP-I
1.5
phosphate
-
pH 7, 30C, SMP-I
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000011
phosphorylated fragment 654-880 of human myosin phosphatase targeting subunit GST-fusion
Sus scrofa
-
-
-
0.00006
phosphorylated fragment 697-880 of human myosin phosphatase targeting subunit GST-fusion
Sus scrofa
-
-
-
0.0014
unphosphorylated fragment 654-880 of human myosin phosphatase targeting subunit GST-fusion
Sus scrofa
-
-
-
0.0061
unphosphorylated fragment 697-880 of human myosin phosphatase targeting subunit GST-fusion
Sus scrofa
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00085
-
in virgin rats
0.00102
-
in pregnant rats with hypertension
0.00121
-
in pregnant rats
0.028
-
purified enzyme, pH 7.6, temperature not specified in the publication
1.6
-
pH 7, 30C, myosin light-chain, enzyme SMP-IV
1.84
-
pH 7, 30C, heavy meromyosin, enzyme SMP-IV
4 - 7.7
-
pH 7, 30C, myosin light-chain, enzyme SMP-I
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
endothelial cells, subunit MYPT1
Manually annotated by BRENDA team
-
smooth muscle
Manually annotated by BRENDA team
-
imaginal disc epithelium of Drosophila
Manually annotated by BRENDA team
-
normal and pulmonary hypertensive fetal sheep, pulmonary artery, aorta and vena cava
Manually annotated by BRENDA team
-
embryonic, large myosin phosphatase target subunit MYPT
Manually annotated by BRENDA team
-
lung pulmonary artery, enzyme specific activity is related to Rho-kinase activation during lung morphogenesis
Manually annotated by BRENDA team
-
coronary artery smooth muscle
Manually annotated by BRENDA team
-
fast and slow muscle fibres
Manually annotated by BRENDA team
-
primary culture of neuronal cells
Manually annotated by BRENDA team
-
from venous blood, myosin phosphatase in platelets is composed of the catalytic subunit of PP-1 plus a subunit similar to the 130 kDa regulatory myosin-binding subunit of the smooth muscle phosphatase, major phosphatase is PP-1, also PP-2A
Manually annotated by BRENDA team
-
phasic smooth muscle
Manually annotated by BRENDA team
-
fetal, MLCP activity in pulmonary hypertensive fetal sheep is significantly lower than in normal fetal sheep
Manually annotated by BRENDA team
-
rat embryonic fibroblast cells, mitotic, 133 kDa myosin-binding subunit
Manually annotated by BRENDA team
-
artery of
Manually annotated by BRENDA team
-
myosin-bound PP-1M
Manually annotated by BRENDA team
-
artery
Manually annotated by BRENDA team
-
phasic smooth muscle
Manually annotated by BRENDA team
-
mesenteric vein, high expression of enzyme with predominance of 3-exon-excluded, leucine-zipper negative MYPT 1 isoform
Manually annotated by BRENDA team
-
fetal, same MLCP activities in normal and pulmonary hypertensive fetal sheep
Manually annotated by BRENDA team
additional information
expression profiles of the heart-specific small subunit hHS-M21 and of the large myosin-binding subunit of MLCP
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
subunit MYPT1, in confluent endothelial cells, at the cell membrane and cell-cell contacts
-
Manually annotated by BRENDA team
-
depending on fixation method for immunohistochemistry, localization of myosin phosphatase target subunit found in nucleus, nucleolus, or cytoplasm
Manually annotated by BRENDA team
-
platelet, only PP-1, not PP-2A
Manually annotated by BRENDA team
-
enzyme may interact with membranes, phosphorylation by protein kinase A may modify interaction, distribution of the 130/133 kDa myosin-binding subunit at the membrane
Manually annotated by BRENDA team
-
depending on fixation method for immunohistochemistry, localization of myosin phosphatase target subunit found in nucleus, nucleolus, or cytoplasm
Manually annotated by BRENDA team
-
133 kDa myosin-binding subunit
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
18500
-
small subunit of MP, mass spectrometry
50000
-
gel filtration
65000
-
catalytic subunit of KAMPPase, gel filtration
110000
-
isoenzyme PP-1M, gel filtration
130000
150000
-
kinase- and myosin-associated protein phosphatase KAMPPase, gel filtration
165000
-
sedimentation equilibrium centrifugation
230000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotrimer
monomer
-
1 * 43000, enzyme SMP-II
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
determination of three-dimensional structure of the 57-residue peptide corresponding to residue 658-714 of myosin phosphatase targeting subunit 1 using multi-dimensional NMR techniques
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, enzyme SMP-I, 20 mM KCl, 20 mM Tris-HCl, pH 7.4, 50% glycerol, 0.5 mM EGTA, 0.5 mM EDTA, 1 mM dithiothreitol, at least 1 year, stable
-
-20C, enzyme SMP-IV, 20 mM KCl, 20 mM Tris-HCl, pH 7.4, 50% glycerol, 0.1 mM EGTA, 0.1 mM EDTA, 1 mM dithiothreitol, 0.1 mM phenylmethylsulfonyl fluoride, several months, stable
-
-20C, several months, stable
-
-70C, enzyme SMP-I, 1 M KCl, 20 mM Tris-HCl, pH 7.4, 0.5 mM EGTA, 0.5 mM EDTA, 1 mM dithiothreitol, at least 1 year, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by immunoprecipitation from primary thymocytes using anti-PP1a, anti-PP1b, anti-MYPT1 antibodies in combination with protein A-Sepharose slurry, myosin phosphatase subunits PP1b and MYPT1 coimmunoprecipitate with HDAC7
-
Flag MYPT1 using anti-Flag affinity chromatography, His tagged proteins by Ni-NTA chromatography
-
GSH affinity chromatography
-
immobilized metal ion affinity chromatography (Ni2+)
-
KAMPPase, copurification of MLCPase and myosin light chain kinase activities, forming a multienzyme complex
-
native enzyme 340fold by ion exchange and hydrophobic interaction chromatography
-
native holoenzyme; recombinant 133 kDa myosin-binding subunit and 20 kDa regulatory subunit, expressed in Escherichia coli
-
native holoenzyme; recombinant 18 kDa small subunit, expressed in Escherichia coli
-
Ni2+-NTA-agarose column chromatography
-
of the native and recombinant proteins by immunoprecipitation
-
of the recombinant peptides by chitin beads
-
recombinant 130 kDa regulatory myosin-binding subunit, expressed in SF9 cells
-
recombinant heart-specific small regulatory subunit isoforms hHS-M21 A and B of MLCP
recombinant myosin-binding subunit of MLCP, expressed in SF9 cells
-
SMP-I, holoenzyme and 38 kDa catalytic subunit
-
SMP-IV, 2000-6133fold
-
SMPP-1M, subunits M110, M21 and PP1C catalytic subunit
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 72.5 kDa N-terminal fragment of the 110 kDa subunit is cloned
-
an 89.6 kDa N-terminal fragment of MYPT1, an isoform of the 130 kDa regulatory targeting subunit of myosin phosphatase, is cloned and sequenced, MYPT is expressed by 2 genes, the two gene classes are termed MYPT1 and MYPT2
cDNAs encoding the 133 kDa myosin-binding subunit and the 20 kDa regulatory subunit are cloned and expressed in Escherichia coli BL21(DE3)
-
cDNAs encoding the 2 isoforms of the 130 kDa myosin binding subunit are cloned and sequenced
-
cDNAs encoding the heart-specific small regulatory subunit isoforms hHS-M21 A and B of MLCP are cloned, sequenced and expressed in Escherichia coli, the small subunit is the product of the same gene as the large subunit: MYPT2, genomic structure of the gene for hHS-M21, the genes MYPT1 and MYPT2 encoding the large myosin-binding subunit MBS are cloned and expressed in Escherichia coli, genomic organisation of the MYPT2 gene, located on chromosome 1q32
cloning of the cDNAs encoding the 130 kDa regulatory myosin-binding subunit MLCPa, the 38 kDa catalytic subunit MLCPc and the 20 kDa regulatory subunit MLCPsr, the full-length 130 kDa subunit is expressed in SF9 insect cells using the baculovirus expression system and sequenced
-
co-expression of each of two myc-tagged myosin PPase targeting subunit 1 (MYPT1) isozymes with HA-tagged MLC phosphatase catalytic subunit CS1beta in COS-7 cells, and co-expression with ERM proteins ezrin, radixin, and moesin in COS-7 cells
-
expressed in Escherichia coli XL-10 Gold cells
-
expressed in Mus musculus
-
expression in DO11/10 T cells, downregulation of myosin phosphatase genes by using siRNA for PP1 isoforms and MYPT1
-
expression of FLAG-tagged full-length catalytic subunit MYPT1 in COS-7 cells, co-expression with Rho-associated kinase
-
expression of wild-type and mutant enzymes in CHO cells using a retroviral vector
-
His-tagged 57-residue peptide corresponding to residue 658-714 of myosin phosphatase targeting subunit 1 expressed in Escherichia coli
-
in Escherichia coli, expression of various C-terminal MYPT1 peptides bearing various combinations of a predicted coiled coil region, extensions preceding this coiled coil region, and the leucine-zipper motif
-
MLCP complex (MYPT1-PP1 dimer) is transiently expressed in COS1 cells
-
myosin-binding subunit of MLCP is cloned and expressed in SF9 cells
-
of MYPT1 in yeast AH109 strain for yeast two hybrid screening, MLCP holoenzyme in Sf9 cells by coinfection of rat His MYPT1, Flag MYPT1, rat PP1delta, and rat M21 expressing viruses, transfection of COS7 or NIH3T3 cells
-
overexpression of MYPT1 and MYPT2 in primary cultures of cardiac myocytes and in COS7 cells using baculoviral and adenoviral expression systems
-
several isoforms of the large targeting subunit MYPT are encoded by 2 genes: MYPT1 and MYPT2, located on chromosomes 12 and 1, structure of MYPT1, MYPT2 is cloned from brain
-
subunits are expressed as a GST-fusion protein in Escherichia coli
-
targeting subunit of myosin phosphatase
-
the 18.5 kDa isoform of the small subunit of MP is cloned and expressed in Escherichia coli KS1000(DE3)
-
the lethal mutation FP41 is a missense mutation changing one of the six highly conserved residues in the catalytic domain, D222N in flw-PA and D91N in flw-PB, it is mapped by meiotic recombination with visible recessive markers to the region between cut (7B4) and vermilion (9F11) and between P-elements PBac(WH)CG34408f03664 (9B7) and PBac(WH)Neb-cGPf02352 (9C4), sequence determination of the wild-type and mutant gene from genomic DNA from single FP41/Y larvae and controls
-
various parts of myosin phosphatase targeting subunit GST-fusions expressed in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
stability of subunits PP1cbeta and MYPT1 is interdependent, knocking down one of the subunits decreases the expression level of the other
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A233S
-
site-directed mutagenesis of subunit PP1cgamma
A281G
-
site-directed mutagenesis of subunit PP1cgamma
DELTA495-1030
-
using a deletion mutant it is shown that the C-terminal domain of human MYPT1(495-1030) is responsible for the binding to the N-terminal portion of myosin light meromyosin
F38A
-
site-directed mutagenesis of subunit MYPT1, the mutant subunit is unable to bind to subunit PP1cbeta
G280A
-
site-directed mutagenesis of subunit PP1cbeta, the mutation does not affect interaction with MYPT1
H237N/K238R
-
site-directed mutagenesis of subunit PP1cgamma, the mutant subunit shows over 90% reduced ability to bind to subunit MYPT1
N124A
-
catalytic subunit mutant, catalytically inactive
N236H/R237K
-
site-directed mutagenesis of subunit PP1cbeta, the mutation does not affect interaction with MYPT1
Q198T
-
site-directed mutagenesis of subunit PP1cgamma, mutant subunit is unable to bind to subunit MYPT1
S232A
-
site-directed mutagenesis of subunit PP1cbeta, the mutation does not affect interaction with MYPT1
T197Q
-
site-directed mutagenesis of subunit PP1cbeta, the mutant subunit is unable to bind to subunit MYPT1
T696A
-
substitution of Thr696 with Ala eliminates the phosphorylation-dependent inhibition
T696D
-
part of myosin phosphatase targeting subunit, inhibitory site
T853A
-
the extent of inhibition of mutant T853A MLCP is indistinguishable from that of wild-type
D884A
-
mutant is not cleaved by caspase-3 in apoptotic cells
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
additional information