Information on EC 3.4.24.65 - macrophage elastase

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The expected taxonomic range for this enzyme is: Euarchontoglires

EC NUMBER
COMMENTARY hide
3.4.24.65
-
RECOMMENDED NAME
GeneOntology No.
macrophage elastase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of soluble and insoluble elastin. Specific cleavages are also produced at -Ala14-/-Leu- and -Tyr16-/-Leu- in the B chain of insulin
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
146888-86-0
-
150680-47-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Mus musculus C57BL/6
C57BL/6 mice
-
-
Manually annotated by BRENDA team
male Wistar rats
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Ala-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Arg-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Glu-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Leu-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Lys-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Phe-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Ser-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Trp-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
2,4-dinitrophenyl-Arg-Pro-Leu-Ala-Tyr-Trp-Arg-Ser-NH2 + H2O
?
show the reaction diagram
-
-
-
?
alpha-1-antitrypsin + H2O
?
show the reaction diagram
-
recombinant enzyme rHME
-
?
alpha-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alpha1-antitrypsin + H2O
?
show the reaction diagram
beta-casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
chondroitan sulfate + H2O
?
show the reaction diagram
-
recombinant enzyme rHME
-
?
Collagen type I + H2O
?
show the reaction diagram
-
the potential of MMP-12 in recognizing sites in human skin collagen types I and III has been investigated. The catalytic domain of MMP-12 binds to the triple helix and cleaves the typical sites -Gly775-Leu776- in alpha-2 type I collagen and -Gly775-Ile776- in alpha-1 type I and type III collagens and at multiple other sites in both collagen types. The region around these typical sites contains comparatively less prolines, of which some have been proven to be only partially hydroxylated
-
-
?
Collagen type III + H2O
?
show the reaction diagram
-
the potential of MMP-12 in recognizing sites in human skin collagen types I and III has been investigated. The catalytic domain of MMP-12 binds to the triple helix and cleaves the typical sites -Gly775-Leu776- in alpha-2 type I collagen and -Gly775-Ile776- in alpha-1 type I and type III collagens and at multiple other sites in both collagen types. The region around these typical sites contains comparatively less prolines, of which some have been proven to be only partially hydroxylated
-
-
?
DQ-collagen I + H2O
?
show the reaction diagram
-
-
-
-
?
DQ-collagen IV + H2O
?
show the reaction diagram
-
-
-
-
?
Elastin + H2O
?
show the reaction diagram
elastin fELN-125 + H2O
?
show the reaction diagram
-
-
-
-
?
enactin + H2O
?
show the reaction diagram
-
recombinant enzyme rHME
-
?
extracellular matrix protein + H2O
?
show the reaction diagram
-
may be required for macrophages to penetrate basement membranes and remodel injured tissue during inflammation
-
?
fEln-100 + H2O
?
show the reaction diagram
-
alpha-elastin
-
-
?
Fibronectin + H2O
?
show the reaction diagram
Gelatin + H2O
?
show the reaction diagram
heparan sulfate + H2O
?
show the reaction diagram
-
recombinant enzyme rHME
-
?
human apolipoprotein (alpha) + H2O
?
show the reaction diagram
-
cleaves in the linker region between kringles IV-4 and IV-5
-
?
Insulin B-chain + H2O
?
show the reaction diagram
-
hydrolyzes Ala-Leu and Tyr-Leu, not His10-Leu11, on amino side of Leu-residue, not Val-residue
-
-
-
Laminin + H2O
?
show the reaction diagram
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 + H2O
?
show the reaction diagram
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 + H2O
Mca-Pro-Leu-Gly + Leu-Dpa-Ala-Arg-NH2
show the reaction diagram
-
-
-
-
?
Mca-Pro-Leu-Gly-Leu-Glu-Glu-Ala-Dpa-NH2 + H2O
Mca-Pro-Leu-Gly + Leu-Glu-Glu-Ala-Dpa-NH2
show the reaction diagram
-
selective cleavage, the Glu-Glu motif interacts with the S'2 and S'3 subsites of MMP-12
-
-
?
TNF-alpha + H2O
?
show the reaction diagram
triple helical peptide alpha1(V) + H2O
?
show the reaction diagram
-
collagen V fibrils
-
-
?
Type IV collagen + H2O
?
show the reaction diagram
type-IV collagen + H2O
?
show the reaction diagram
-
degradation
-
-
?
Elastin + H2O
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
casein + H2O
?
show the reaction diagram
-
degradation
-
-
?
Elastin + H2O
?
show the reaction diagram
extracellular matrix protein + H2O
?
show the reaction diagram
-
may be required for macrophages to penetrate basement membranes and remodel injured tissue during inflammation
-
?
Fibronectin + H2O
?
show the reaction diagram
-
degradation
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Laminin + H2O
?
show the reaction diagram
-
degradation
-
-
?
type-IV collagen + H2O
?
show the reaction diagram
-
degradation
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1S,5S,7R)-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-3,7-dicarboxylic acid 7-[(biphenyl-4-ylmethyl)-amide] 3-hydroxyamide
-
-
(1S,5S,7R)-3-biphenyl-4-ylmethyl-2-oxo-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-7-carboxylic acid
-
-
(1S,5S,7R)-3-biphenyl-4-ylmethyl-2-oxo-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-7-carboxylic acid hydroxyamide
-
-
(1S,5S,7R)-3-biphenyl-4-ylmethyl-3-aza-6,8-dioxabicyclo[3.2.1]octane-7-carboxylic acid
-
-
(1S,5S,7R)-3-biphenyl-4-ylmethyl-3-aza-6,8-dioxabicyclo[3.2.1]octane-7-carboxylic acid hydroxyamide
-
-
(3R)-3-([[4-(4'-acetylbiphenyl-4-yl)-5-fluorothiophen-2-yl]carbonyl]amino)-3-phenylpropanoic acid
-
-
(3R)-3-[([5-fluoro-4-[4-(pyridin-4-yl)phenyl]thiophen-2-yl]carbonyl)amino]-3-phenylpropanoic acid
-
-
(3R)-3-[([5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]-3-phenylpropanoic acid
-
-
alpha2-Macroglobulin
batimastat
BB-94
CGS-27023A
-
a hydroxamate inhibitor
CGS27023A
-
-
dexamethasone
-
in vivo inhibition of the human enzyme in mice leading to inhibited cytokine release and neutrophil influx, overview
dithiothreitol
-
-
EGTA
-
less effective than EDTA, Ca2+ at a 3:1 ratio of Ca2+/EDTA protects and reverses partially, Zn2+ at a 1:10 ratio of Zn2+/EGTA protects and reverses
marimastat
-
in vivo inhibition of the human enzyme in mice leading to inhibited macrophage recruitment, neutrophil influx, and cytokine release, overview
N-(3-[(4-bromophenyl)(hydroxy)phosphoryl]-2-[3-[4-(dimethylamino)phenyl]isoxazol-5-yl]propanoyl)-L-alpha-glutamyl-L-alpha-glutamine
-
-
N-isobutyl-N-[4-methoxyphenylsulfonyl] glycyl hydroxamic acid
-
-
N-[(1R)-3-amino-3-oxo-1-phenylpropyl]-5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide
-
-
N-[3-[(4-bromophenyl)(hydroxy)phosphoryl]-2-[3-(3'-chlorobiphenyl-4-yl)isoxazol-5-yl]propanoyl]-L-alpha-glutamyl-L-alpha-glutamine
-
-
Organic solvents
-
enzyme form B, above 0.5% v/v
-
Rabbit antiserum against purified elastase form B
-
-
-
rolipram
-
in vivo inhibition of the human enzyme in mice leading to inhibited neutrophil influx, overview
TIMP-1
-
a major fibrogenic effector in lungs. Indeed, upon fibrogenic stimuli, large amounts of TIMP-1 in the remodeling tissue are believed to contribute to the creation of a non-degrading environment, leading to the alteration of protease-anti-protease balance, extracellular matrix accumulation, and tissue fibrosis. MMP-12 deficiency does not seem to be involved in TIMP-1 regulation
-
Tissue inhibitor of metalloproteinase
trans-4-[([[4-(4'-acetylbiphenyl-4-yl)-5-fluorothiophen-2-yl]carbonyl]amino)methyl]cyclohexanecarboxylic acid
-
-
trans-4-[[([3-[4-(trifluoromethoxy)phenyl]-1,2-thiazol-5-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
-
-
trans-4-[[([4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
-
-
trans-4-[[([5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
-
-
trans-4-[[([5-[4-(trifluoromethoxy)phenyl]-1,2-thiazol-3-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
-
-
Zn2+
-
at a 3fold molar excess of Zn2+ over chelating agent
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 0.00466
fEln-100
0.113 - 0.241
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 0.021
fEln-100
6.8 - 24.1
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
34.6
DQ-collagen I
Homo sapiens
-
pH 7.5, 25C
168024
29.2
DQ-collagen IV
Homo sapiens
-
pH 7.5, 25C
161170
9.8
elastin fELN-125
Homo sapiens
-
pH 7.5, 25C
168023
0.59 - 10.69
fEln-100
51.57 - 159.7
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
55.4
triple helical peptide alpha1(V)
Homo sapiens
-
pH 7.5, 25C
168025
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000044
N-(3-[(4-bromophenyl)(hydroxy)phosphoryl]-2-[3-[4-(dimethylamino)phenyl]isoxazol-5-yl]propanoyl)-L-alpha-glutamyl-L-alpha-glutamine
-
-
0.00000019
N-[3-[(4-bromophenyl)(hydroxy)phosphoryl]-2-[3-(3'-chlorobiphenyl-4-yl)isoxazol-5-yl]propanoyl]-L-alpha-glutamyl-L-alpha-glutamine
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
149
(1S,5S,7R)-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-3,7-dicarboxylic acid 7-[(biphenyl-4-ylmethyl)-amide] 3-hydroxyamide
Homo sapiens
-
pH 7.0, 25C, recombinant enzyme
0.954
(1S,5S,7R)-3-biphenyl-4-ylmethyl-2-oxo-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-7-carboxylic acid
Homo sapiens
-
pH 7.0, 25C, recombinant enzyme
425
(1S,5S,7R)-3-biphenyl-4-ylmethyl-2-oxo-3-aza-6,8-dioxa-bicyclo[3.2.1]octane-7-carboxylic acid hydroxyamide
Homo sapiens
-
pH 7.0, 25C, recombinant enzyme
835
(1S,5S,7R)-3-biphenyl-4-ylmethyl-3-aza-6,8-dioxabicyclo[3.2.1]octane-7-carboxylic acid
Homo sapiens
-
pH 7.0, 25C, recombinant enzyme
399
(1S,5S,7R)-3-biphenyl-4-ylmethyl-3-aza-6,8-dioxabicyclo[3.2.1]octane-7-carboxylic acid hydroxyamide
Homo sapiens
-
pH 7.0, 25C, recombinant enzyme
0.000024
(3R)-3-([[4-(4'-acetylbiphenyl-4-yl)-5-fluorothiophen-2-yl]carbonyl]amino)-3-phenylpropanoic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.000013
(3R)-3-[([5-fluoro-4-[4-(pyridin-4-yl)phenyl]thiophen-2-yl]carbonyl)amino]-3-phenylpropanoic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.0002
(3R)-3-[([5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]-3-phenylpropanoic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.000085
N-[(1R)-3-amino-3-oxo-1-phenylpropyl]-5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide
Homo sapiens
-
pH and temperature not specified in the publication
0.000007
trans-4-[([[4-(4'-acetylbiphenyl-4-yl)-5-fluorothiophen-2-yl]carbonyl]amino)methyl]cyclohexanecarboxylic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.0036
trans-4-[[([3-[4-(trifluoromethoxy)phenyl]-1,2-thiazol-5-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.0004
trans-4-[[([4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.00014
trans-4-[[([5-fluoro-4-[4-(trifluoromethoxy)phenyl]thiophen-2-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.031
trans-4-[[([5-[4-(trifluoromethoxy)phenyl]-1,2-thiazol-3-yl]carbonyl)amino]methyl]cyclohexanecarboxylic acid
Homo sapiens
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
-
assay at
7
-
assay at
8.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
33
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
MMP-12/tissue inhibitor of metalloproteinase-1 ratio is significantly increased in the CSF of Angiostrongylus cantonensis-infected mice from day 10 p.i., and reaches high levels on days 20 and 25 p.i. MMP-12 production is correlated with elastin degradation, eosinophil count, blood-CSF barrier permeability and pathological changes in the subarachnoid space
Manually annotated by BRENDA team
-
bronchial
Manually annotated by BRENDA team
-
HME protein levels are 4.50-7.15fold increased in gastric carcinoma samples compared with the non-cancerous controls
Manually annotated by BRENDA team
-
irregular glomerular basement membrane that characterizes Alport syndrome may be mediated, in part, by focal degradation of glomerular basement membrane due to MMP dysregulation, in particular, MMP-12
Manually annotated by BRENDA team
-
only in glomerulonephritic ICGN strain, not in non-glomerulonephritic mouse (immunohistochemistry)
Manually annotated by BRENDA team
-
macrophage-like
Manually annotated by BRENDA team
-
ICGN strain, small amounts during early pogression of renal disease, increasing amounts with progressing renal disease (immunohistochemistry)
Manually annotated by BRENDA team
-
16-50 SCCs, in situ detection, overview
Manually annotated by BRENDA team
-
HME is expressed in chronic gastritis with atypical hyperplasia and normal gastric epithelium mucosa. HME protein levels are 4.50-7.15fold increased in gastric carcinoma samples compared with the non-cancerous controls
Manually annotated by BRENDA team
-
mainly expressed by inflammatory macrophages in the rhematoid arthritis synovial membrane. Rheumatoid synovial tissue contains higher levels of MMP-12 messenger RNA than does osteoarthritis synovial tissue. Macrophage-derived MMP-12 may play an important role in the destructive process in rheumatoid arthritis
Manually annotated by BRENDA team
additional information
-
immunohistochemical expression analysis
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
recombinant enzyme from Escherichia coli
Manually annotated by BRENDA team
-
glomerular basement membrane. Irregular glomerular basement membrane that characterizes Alport syndrome may be mediated, in part, by focal degradation of glomerular basement membrane due to MMP dysregulation, in particular, MMP-12
Manually annotated by BRENDA team
additional information
-
MMP12, in contrast to other MMPs, can act intracellularly rather than extracellularly. The majority of MMP12 is secreted, MMP12 might then bind to the bacteria outside the cell prior to phagocytosis
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22000
-
mouse, form B and C, gel filtration, sucrose density gradient centrifugation
45000
-
MMp-12 active enzyme
54000
-
MMp-12 proenzyme
57000
-
mouse, form A, sucrose density gradient centrifugation, gel filtration
59000
-
SDS-PAGE
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 22000, mouse, form B and C, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
-
-
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hexagonal shaped crystals, crystal structure in complex with hydroxamic acid inhibitor, orthorhombic space group I222, unit cells dimensions a : 67.4 A, b : 87.2, c : 169.2
-
purified MMP-12 bound to hydroxamic acid, hanging drop vapour diffusion method, 20C, from protein solution containing 10 mg/ml protein in 0.1 M Tris-HCl, pH 8.0, 30% PEG 6000, 200 mM hydroxamic acid, and 1.0 M LiCl, a few days, X-ray diffraction structure determination and analysis at 1.3 A resolution
-
sitting drop vapor diffusion technique, crystallized in complex with inhibitor batimastat, BB-94, monoclinic space group C2, crystal cell constants a : 51.91 A, b : 60.26 A, c : 54.61 A
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Buffers with high concentration of Ca2+ stabilize during purification
-
catalytic domain of MMP-12 exhibits higher activity, more rigidity of its backbone, and lower folding stability than its counterpart of the MMP-3 catalytic domain that has more internal motions throughout
-
Dialysis against 25 mM Tris-HCl, pH 7.6 inactivates, Ca2+ restores activity
-
Purification in the presence of EDTA prevents activation of proenzyme
-
Purified enzyme, not partially purified preparation, unstable in either glycerol or sucrose velocity gradients
-
Stable to freeze-thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, in 50 mM Tris-HCl, pH 7.6, 150 mM NaCl, 50 mM CaCl2, at least 3 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 chromatographically distinct forms A, B and C, predominant form is B
-
partial, recombinant or native enzyme
-
recombinant F171D mutant fragment Gly106-Gly263 from Escherichia coli strain BL21 by gel filtration and anion exchange chromatography
-
recombinant MMP-12
-
refolded recombinant MMP-12 catalytic domain from Escherichia coli strain BL21(DE3) inclusion bodies by anion exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned and expressed in Escherichia coli BL21(DE3)
-
expressed in Escherichia coli
expression analysis
-
expression analysis of MMP12 in cancerous and healthy oral tissues, overview
-
expression of a cDNA encoding the fragment Gly106-Gly263 of the enzyme F171D mutant in Escherichia coli strain BL21
-
generation of transgenic rabbits that express human (h)MMP-12 gene under the control of a macrophage-specific promoter, the human scavenger receptor promoter. This transgenic rabbit model with increased expression of hMMP-12 may become a useful model for further mechanistic studies of MMP-12 in in inflammatory diseases and cancer invasion, it is also an ideal model for testing the in vivo action of MMP-12 inhibitors
-
MMP-12 catalytic domain, cd(EA)MMP-12 overexpressed in Escherichia coli strain BL21 (DE3)
MMP-12 mRNA and protein level increased with the progression of renal disease in glomerulonephritic ICGN mouse compared to non-glomerulonephritic mouse
-
molecular cloning of the proteinase domain of MMP12
-
overexpression of the MMP-12 catalytic domain, residues Phe100-Gly263, in Escherichia coli strain BL21(DE3) inclusion bodies
-
stable expression of MME domains 1 and 2 in enzyme-deficient murine CT-26 colon cancer cells suppresses orthotopic tumor growth, angiogenesis and vascular endothelial growth factor expression, expression analysis and orthotopic animal tumor model study, overview
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
MMP-12 expression is upregulated by other matrix components such as hyaluronan fragments, cytokines, and growth factors, for example by TGF-beta, IFN-gamma and EGF, and serine proteases such as thrombin and plasmin
MMP-12/tissue inhibitor of metalloproteinase-1 ratio is significantly increased in the CSF of Angiostrongylus cantonensis-infected mice from day 10 p.i., and reaches high levels on days 20 and 25 p.i. MMP-12 production is correlated with elastin degradation, eosinophil count, blood-CSF barrier permeability and pathological changes in the subarachnoid space
-
UV radiation is known to induce the expression of MMP-12 in skin
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A164V
-
mutation inconsequential to elastolysis
A182G
-
mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows strong decreased (kcat/Km) toward fEln-100 compared to wild-type
D124Q
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mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows decreased (kcat/Km) toward fEln-100 compared to wild-type
D124Q/A182G
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combination of two well-separated mutations further reduces activity toward both fEln-100 and elastin-fluorescein compared to the single mutations, mutant shows decreased activity towards MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
D124Q/I180S
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combination of two well-separated mutations further reduces activity toward both fEln-100 and elastin-fluorescein compared to the single mutations, mutant shows decreased activity towards MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
D124Q/M156E
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combination of two well-separated mutations further reduces activity toward both fEln-100 and elastin-fluorescein compared to the single mutations, mutant retains wild-type activity towards MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
D124Q/M156E/A182G
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mutant confers a significantly greater loss of catalytic efficiency in digesting fEln-100 than the parental double mutant D124Q/M156E, catalytic activity toward MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 is highly decreased compared to wild-type, compared to triple mutant D124Q/M156E/F185Y and D124Q/M156E/T205K catalytic efficacy toward fEln-100 is highly decreased
D124Q/M156E/F185Y
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mutant confers a significantly greater loss of catalytic efficiency in digesting fEln-100 than the parental double mutant D124Q/M156E, catalytic activity toward MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 is not affected compared to wild-type, compared to triple mutant D124Q/M156E/I180S and D124Q/M156E/A182G catalytic efficacy toward fEln-100 is moderately decreased
D124Q/M156E/I180S
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mutant confers a significantly greater loss of catalytic efficiency in digesting fEln-100 than the parental double mutant D124Q/M156E, catalytic activity toward MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 is highly decreased compared to wild-type, compared to triple mutant D124Q/M156E/F185Y and D124Q/M156E/T205K catalytic efficacy toward fEln-100 is highly decreased
D124Q/M156E/T205K
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mutant confers a significantly greater loss of catalytic efficiency in digesting fEln-100 than the parental double mutant D124Q/M156E, catalytic activity toward MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 is not affected compared to wild-type, compared to triple mutant D124Q/M156E/I180S and D124Q/M156E/A182G catalytic efficacy toward fEln-100 is moderately decreased
D200E
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mutation inconsequential to elastolysis
G166R
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mutation inconsequential to elastolysis
I180S
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mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows strong decreased (kcat/Km) toward fEln-100 compared to wild-type
I255V
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mutation inconsequential to elastolysis
K148T
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mutation inconsequential to elastolysis
M103F
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mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows decreased catalytic efficacy (kcat/Km) toward fEln-100 compared to wild-type
M156E
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mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows strong decreased (kcat/Km) toward fEln-100 compared to wild-type
M156E/A182G
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kcat (fEln-100) is twice that of wild-type, mutant retains wild-type activity towards MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
M156E/I180S
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combination of two well-separated mutations further reduces activity toward both fEln-100 and elastin-fluorescein compared to the single mutations, mutant shows decreased activity towards MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
N153Y
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mutation inconsequential to elastolysis
R117S
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mutant retains similar activity toward substrate MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Km and kcat similar to wild-type, kcat/Km), mutant shows decreased (kcat/Km) toward fEln-100 compared to wild-type
S142E
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mutation inconsequential to elastolysis
V144A
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mutation inconsequential to elastolysis
V162S
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mutation inconsequential to elastolysis
Y132A
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mutation inconsequential to elastolysis
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant fragment of mutant F171D from Escherichia coli inclusion bodies by solubilization in 20 mM Tris-HCl and 8 M urea at pH 8.0, refolding by multi-step dialysis, overview
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recombinant MMP-12 catalytic domain from Escherichia coli strain BL21(DE3) inclusion bodies with refolding buffer, which contains 6 M urea, 20 mM Tris-HCl, 5 mM CaCl2, and 100 mM NaCl, pH 7.5, followed by a two-step dialysis against refolding buffer containing 0.1 mM ZnCl2, overview
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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the enzyme can be utilized for pharmacological evaluation of anti-inflammatory mechanisms of action
diagnostics
drug development
medicine
pharmacology