Information on EC 1.14.11.4 - procollagen-lysine 5-dioxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.11.4
-
RECOMMENDED NAME
GeneOntology No.
procollagen-lysine 5-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-lysine-[procollagen] + 2-oxoglutarate + O2 = (2S,5R)- 5-hydroxy-L-lysine-[procollagen] + succinate + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
-
-
-
-
hydroxylation
redox reaction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
procollagen hydroxylation and glycosylation
-
-
Lysine degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
L-lysine-[procollagen],2-oxoglutarate:oxygen oxidoreductase (5-hydroxylating)
Requires Fe2+ and ascorbate.
CAS REGISTRY NUMBER
COMMENTARY hide
9059-25-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
zebrafish
A0JMD1, A4U7F8, A4U7F9
TrEMBL
Manually annotated by BRENDA team
gene dPlod, or CG6199, encoding the only lysyl hydroxylase of the organism
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
gene Plod3
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
-
heterocomplex formation between mouse and human LH3, between human LH1 and LH3 and between human LH2 and LH3, low amount of complexes formed in vivo
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(GDK)4 + 2-oxoglutarate + O2
?
show the reaction diagram
(IKG)3 + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
(Ile-Lys-Gly)3 + 2-oxoglutarate + O2
(Ile-5-hydroxylyseine-Gly)3 + succinate + CO2
show the reaction diagram
(Ile-Lys-Gly)3 + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
(Pro-Pro-Gly)4-Ala-Arg-Gly-Met-Lys-Gly-His-Arg-Gly-(Pro-Pro-Gly)4 + 2-oxoglutarate + O2
(Pro-Pro-Gly)4-Ala-Arg-Gly-Met-5-hydroxylysine-Gly-His-Arg-Gly-(Pro-Pro-Gly)4 + succinate + CO2
show the reaction diagram
-
-
-
?
2-oxoglutarate + O2 + ascorbate
succinate + CO2 + dehydroascorbate + H2O
show the reaction diagram
adiponectin-L-lysine + 2-oxoglutarate + O2
adiponectin-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
Ala-Arg-Gly-Ile-Lys-Gly-Ile-Arg-Gly-Phe-Ser-Gly + 2-oxoglutarate + O2
Ala-Arg-Gly-Ile-5-hydroxylysine-Gly-Ile-Arg-Gly-Phe-Ser-Gly + succinate + CO2
show the reaction diagram
Ala-Arg-Gly-Met-Lys-Gly-His-Arg-Gly-(Pro-Pro-Gly)4 + 2-oxoglutarate + O2
Ala-Arg-Gly-Met-5-hydroxylysine-Gly-His-Arg-Gly-(Pro-Pro-Gly)4 + succinate + CO2
show the reaction diagram
-
-
-
?
collagen + 2-oxoglutarate + O2
5-hydroxylysyl-collagen + succinate + CO2
show the reaction diagram
KGIKGIKG + 2-oxoglutarate + O2
?
show the reaction diagram
L-lysine containing nonapeptides + 2-oxoglutarate + O2
5-hydroxy-L-lysine containing nonapeptides + succinate + CO2
show the reaction diagram
-
diverse nonapeptides, synthetic substrates
-
-
?
L-lysine-[collagen] + 2-oxoglutarate + O2
5-hydroxy-L-lysine-[collagen] + succinate + CO2
show the reaction diagram
L-lysine-[procollagen] + 2-oxoglutarate + O2
(2S,5R)-5-hydroxy-L-lysine-[procollagen] + succinate + CO2
show the reaction diagram
L-lysine-[U2AF65] + 2-oxoglutarate + O2
5-hydroxy-L-lysine-[U2AF65] + succinate + CO2
show the reaction diagram
luc7like2(267-278) + 2-oxoglutarate
? + succinate + CO2
show the reaction diagram
substrate is a Luc7like2 protein fragment
-
-
?
mannan-binding lectin-A-L-lysine + 2-oxoglutarate + O2
mannan-binding lectin-A-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
peptide (IKG)3 + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
peptidyl-L-lysine + 2-oxoglutarate + O2
peptidyl-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
procollagen L-lysine + 2-oxoglutarate + O2
procollagen 5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
type I procollagen + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
type IV procollagen + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
U2AF65 + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
[procollagen]-L-lysine + 2-oxoglutarate + O2
[procollagen]-(2S,5R)-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
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
adiponectin-L-lysine + 2-oxoglutarate + O2
adiponectin-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
collagen + 2-oxoglutarate + O2
5-hydroxylysyl-collagen + succinate + CO2
show the reaction diagram
L-lysine-[collagen] + 2-oxoglutarate + O2
5-hydroxy-L-lysine-[collagen] + succinate + CO2
show the reaction diagram
L-lysine-[procollagen] + 2-oxoglutarate + O2
(2S,5R)-5-hydroxy-L-lysine-[procollagen] + succinate + CO2
show the reaction diagram
L-lysine-[U2AF65] + 2-oxoglutarate + O2
5-hydroxy-L-lysine-[U2AF65] + succinate + CO2
show the reaction diagram
-
U2AF65 is the splicing factor U2 small nuclear ribonucleoprotein auxiliary factor 65-kDa subunit, which undergoes posttranslational lysyl-5-hydroxylation catalyzed by the Fe2+ and 2-oxoglutarate-dependent dioxygenase Jumonji domain-6 protein Jmjd6, a nuclear protein that has an important role in vertebrate development and is a human homologue of the HIF asparaginyl-hydroxylase
-
-
?
mannan-binding lectin-A-L-lysine + 2-oxoglutarate + O2
mannan-binding lectin-A-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
procollagen L-lysine + 2-oxoglutarate + O2
procollagen 5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
[procollagen]-L-lysine + 2-oxoglutarate + O2
[procollagen]-(2S,5R)-5-hydroxy-L-lysine + succinate + CO2
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ascorbate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
-
reduces the activity of the isozymes LH1, LH2a, LH2b, and LH3 with most of the synthetic nonpeptide substrates tested, in some cases the isozyme is activated or completely inhibited, overview
Ni2+
can substitute for Fe2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoglutarate
-
competitive substrate inhibition
Adrenochrome
-
slight
catechol
-
-
dehydroascorbate
-
-
dipyridyl
a non-specific hydroxylase inhibitor; a non-specific hydroxylase inhibitor; a non-specific hydroxylase inhibitor
DL-Serine 2-[(2,3,4-trihydroxyphenyl)methyl]hydrazide
-
most potent
dopamine
-
-
ephedrine
-
slight
epinephrine
-
-
homogentisic acid
-
-
Hydroxylysine-rich peptides
-
-
iodoacetamide
-
-
L-alpha-hydroxyglutarate
-
the enzyme activity is inhibited by L-alpha-hydroxyglutarate but not its enantiomer D-alpha-hydroxyglutarate
-
malaoxon
-
mechanism of inhibition
malathion
-
mechanism of inhibition
minoxidil
N-ethylmaleimide
-
-
NaCl
-
reduces the activity of the isozymes LH1, LH2a, LH2b, and LH3 with most of the synthetic nonpeptide substrates tested, in some cases the isozyme is activated or completely inhibited, overview
nitroblue tetrazolium
-
-
norepinephrine
-
-
p-chloromercuribenzoate
-
-
p-mercuribenzoate
-
-
phenylalanine
-
slight
Phenylephedrine
-
slight
-
Pyridine 2,4-dicarboxylate
-
-
Pyridine 2,5-dicarboxylate
-
-
pyridine 2-carboxylate
-
-
pyrogallol
-
most potent
SC65
directly interacts with lysyl-hydroxylase 1, LH1
-
succinate
-
-
tyrosine
-
slight
additional information
the endoplasmic reticulum complex including SC65 and prolyl 3-hydroxylase 3affects the activity of lysyl-hydroxylase 1 potentially through interactions with the enzyme and/or cyclophilin B. Loss of Sc65 in the mouse results in instability of this complex, altered collagen lysine hydroxylation and cross-linking leading to connective tissue defects that include low bone mass and skin fragility, while it has no effect on prolyl 3-hydroxylation. CRTAP, a non-enzymic member of the Leprecan family of proteins which includes Synaptonemal Complex 65 (SC65) and the prolyl 3-hydroxylases (P3H1, P3H2 and P3H3), is an essential third subunit of a complex with prolyl 3-hydroxylase 1 (aka LEPRECAN) and cyclophilin B (CYPB) in the endoplasmic reticulum, forming the so-called collagen prolyl 3-hydroxylation complex
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine
-
can replace ascorbate
2-mercaptoethanol
-
can replace ascorbate
ascorbate
-
required for activity
ascorbic acid
bovine serum albumin
-
cannabidiol
CBD, a major non-psychotropic cannabis constituent, stimulates lysyl hydroxylase 1 activity in osteoblasts, cannabidiol enhances the biomechanical properties of healing rat mid-femoral fractures
catalase
-
DELTA9-tetrahydrocannabinol
THC, the psychotropic cannabis constituent, stimulates lysyl hydroxylase 2 activity in osteoblasts
dithiothreitol
FKBP65
FKBP65 interacts with LH2 through its peptidyl prolyl isomerase (PPIase) domains, FKBP65 forms a complex with LH2. Reconstitution with wild-type FKBP65 increases the enzyme activity by 7fold
-
L-cysteine
-
can replace ascorbate
lysolecithin
-
activation
Triton X-100
-
activation
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4 - 5
(IKG)3
0.31 - 0.8
(Ile-Lys-Gly)3
0.43
(L-Ile-L-Lys-Gly)3
-
lysyl hydroxylase 3
0.2
(Pro-Pro-Gly)4-Ala-Arg-Gly-Met-Lys-Gly-His-Arg-Gly-(Pro-Pro-Gly)4
-
-
0.011 - 0.25
2-oxoglutarate
0.4 - 0.6
Ala-Arg-Gly-Ile-Lys-Gly-Ile-Arg-Gly-Phe-Ser-Gly
0.2
Ala-Arg-Gly-Met-Lys-Gly-His-Arg-Gly-(Pro-Pro-Gly)4
-
-
0.05 - 0.35
ascorbate
0.04 - 0.05
O2
-
-
0.169
peptide (IKG)3
-
pH 7.4, 37°C, recombinant wild-type enzyme
0.1 - 0.5
Protocollagen
-
0.08 - 0.23
type I procollagen
-
0.04 - 0.3
type IV procollagen
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.183
2-oxoglutarate
-
pH 7.4, 37°C, recombinant wild-type enzyme
2.6 - 4.2
lysine
-
in synthetic peptides
0.2
peptide (IKG)3
-
pH 7.4, 37°C, recombinant wild-type enzyme
additional information
additional information
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.6
2-oxoglutarate
-
pH 7.4, 37°C, recombinant wild-type enzyme
1.18
peptide (IKG)3
-
pH 7.4, 37°C, recombinant wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015
catechol
-
-
0.047
malaoxon
-
-
0.059
malathion
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
assay at
8 - 8.4
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8
A5A6S1;, A5A6S2;, A5A6S3;
isozyme LH1, sequence calculation
6.1
A5A6S1;, A5A6S2;, A5A6S3;
isozyme LH3, sequence calculation
6.2
A5A6S1;, A5A6S2;, A5A6S3;
isozyme LH2a, sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
p21-deficient (KC) and -replete (K) murine lung adenocarcinoma cells, increased LH2 enzyme activity
Manually annotated by BRENDA team
expression only of LH2long
Manually annotated by BRENDA team
stem marrow cell, no expression of isozymes LH3 and LH2b
Manually annotated by BRENDA team
expression only of LH2long
Manually annotated by BRENDA team
A5A6S1;, A5A6S2;, A5A6S3;
-
Manually annotated by BRENDA team
A5A6S1;, A5A6S2;, A5A6S3;
-
Manually annotated by BRENDA team
A5A6S1;, A5A6S2;, A5A6S3;
in the hatching larvae, LH1 is expressed in the pectoral fin bud; in the hatching larvae, LH3 is expressed in the pectoral fin bud
Manually annotated by BRENDA team
4 lung adenocarcinoma cell lines (KC1–KC4) from lung tumors in KC mice and 2 lung adenocarcinoma cell lines (K1 and K2) from Cdkn1aWT K-rasLA1 mice. Expression levels of the third gene of interest, LH2, are 10-30 times higher in the highly metastatic KC cells and include both LH2 isoforms, full-length (LH2b) and spliced at exon 13 (LH2a)
Manually annotated by BRENDA team
-
increased LH2 enzyme activity in lung carcinoma compared to healthy lung
Manually annotated by BRENDA team
A5A6S1;, A5A6S2;, A5A6S3;
high expression of lysine hydroxylase 3
Manually annotated by BRENDA team
lysyl hydroxylases are expressed in white-adipose tissue of ob/ob mice, overview; lysyl hydroxylases are expressed in white-adipose tissue of ob/ob mice, wherein LH3 levels are increased compared with C57/BL controls. Expression levels of mRNAs corresponding to LH1 (Plod1) and LH3 (Plod3) are both altered in ob/ob mice, in the same direction as the changes in serum levels of adiponectin and the HMW isoforms; lysyl hydroxylases are expressed in white-adipose tissue of ob/ob mice, wherein LH3 levels are increased compared with C57/BL controls. Expression levels of mRNAs corresponding to LH1 (Plod1) and LH3 (Plod3) are both altered in ob/ob mice, in the same direction as the changes in serum levels of adiponectin and the HMW isoforms
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
UNIPROT
ORGANISM
Acanthamoeba polyphaga mimivirus;
Homo sapiens;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
x * 42000, recombinant His-tagged catalytic domain of JMJD6, residues 1-343
70000
-
x * 70000 + x * 115000, SDS-PAGE
80000
-
value about, Western blot
82380
-
2 * 82380, calculation from amino acid sequence, recombinant enzyme: isoform 3
84385
A5A6S1;, A5A6S2;, A5A6S3;
x * 84385, isozyme LH1, sequence calculation
84558
A5A6S1;, A5A6S2;, A5A6S3;
x * 84558, isozyme LH3, sequence calculation
85063
A5A6S1;, A5A6S2;, A5A6S3;
x * 85063, isozyme LH2a, sequence calculation
88000
-
2 * 88000 or 97000 SDS-PAGE, recombinant enzymes: isoform 2, possibly two forms are due to variation in the glycosylation of enzymes
89000
-
LH2, SDS-PAGE
97000
-
SDS-PAGE, V5-tagged LH2b
115000
-
x * 70000 + x * 115000, SDS-PAGE
150000
-
gel filtration
180000
-
gel filtration, isoforms 1-3
200000
220000
-
gel filtration, peak 1
550000
-
gel filtration, peak 2
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
additional information
structure of the catalytic domain of recombinant His-tagged JMJD6 and active site structure, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
side-chain modification
additional information
-
posttranslational hydroxylation in both the cytoplasm and the nucleoplasm is ubiquitous
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant His-tagged SeMet-derivatised and wild-type JMJD61-343, at both 20°C and 4°C by the sitting drop vapour diffusion method, X-ray diffraction structure determination and analysis at 1.75-2.0 A resolution
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme formes aggregates in low ionic strength buffer
-
inactivation by freezing/thawing
labile in tissue extracts
loss of activity during concentration
stabilization by detergents, NaCl
stabilization by glycine
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C
-20°C, enzyme purified by collagen-agarose column-chromatography stable, enzyme from Bio-gel column unstable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA-agarose
ProBond metal chelate affinity resin chromatography and Chelating Sepharose fast flow column chromatography
-
recombinant His-tagged full-length JMJD6 and JMJD61-343 from Escherichia coli strain Rosetta by nickel affinity chromatography
recombinant His-tagged isozymes LH1-3 from Sf9 insect cells by nickel affinity chromatography
-
recombinant His10-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant V5/His-tagged LH3 protein from HEK-293 cells by nickel affinity chromatography, dialysis, and ultrafiltration
recombinant wild-type and mutant enzymes from Sf9 insect cells by chelating affinity chromatography
-
three recombinant isoenzymes
-
two alternative procedures
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
all 3 isozymes LH1-3, LH2 in 2 splicing variants, are expressed in as His-tagged proteins in Spodoptera frugiperda Sf9 cells via baculovirus infection system with the signal peptide at the N-terminus in isozymes LH2a and b and LH3 being exchanged for the signal peptide of isozyme LH1
-
cloning of 3 isozymes LH1-3, with 2 splicing variants of isozyme LH2, LH2a and LH2b, stable and functional expression in CHO-K1 cells; cloning of 3 isozymes LH1-3, with 2 splicing variants of isozyme LH2, LH2a and LH2b, stable and functional expression in CHO-K1 cells
DNA and amino acid sequence determination and analysis of mutant DNA from an Ehlers-Danlos syndrome type IVA patient, expression of mutant enzyme W446G in an insect cell system via baculovirus infection
-
endogenous LH2 alternate splicing pattern and conservation analysis of LH2 genomic sequence., overview. Construction and expression of a functional LH2 minigene in HEK-293 cells, human neonatal skin fibroblasts, and mouse embryonic fibroblasts. The TIA proteins play a role in the regulation of the alternative splicing of LH2, overview
-
expressed in H5 insect cells
-
expression in Escherichia coli, isoform 3 gene product possesses the collagen glycosyltransferase activity, but not isoform 1 and 2
-
expression in insect cells using a baculovirus vector
expression in insect cells using a baculovirus vector; expression in various human tissues
-
expression in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system, and functional expression of N-terminally His10-tagged enzyme in Escherichia coli strain BL21(DE3)
-
expression of 40 amino acid residues of the C-terminal end of isozyme LH1 fused to human cathepsin D and c-Myc-tagged and 2 mutant variants thereof in COS-7 cells
-
expression of a synthetic gene encoding LH3, SEQ ID, in Nicotiana tabacum leaves. Co-expression of the human genes encoding recombinant heterotrimeric collagen type I, rhCOL1, the human prolyl-4-hydroxylase, and the lysyl hydroxylase 3, all responsible for key posttranslational modifications of collagen. Plants coexpressing all five vacuole-targeted proteins generate intact procollagen yields ofabout 2% of the extracted total soluble protein, overview
-
expression of active LH2b and of antisense construct in MC3T3-E1 cells, the latter suppresses the endogenous enzyme
-
expression of GFP-tagged Jmjd6 in HEK-293 cells, GFP pulldown interaction analysis with U2AF65, overview
-
expression of His-tagged full-length JMJD6 and JMJD61-343 in Escherichia coli strain Rosetta
expression of wild-type and mutantenzymes in Spodoptera frugiperda Sf9 cells using the baculovirus transfection method
-
gene dPlod or CG6199, maps to cytological position 68B1 on the left arm of the third chromosome, DNA and amino acid sequence determination and analysis, sequence comparisons
-
gene encoding isozyme LH1, DNA and amino acid sequence determination and analysis, quantitative expression analysis; gene encoding isozyme LH2a, DNA and amino acid sequence determination and analysis, quantitative expression analysis; gene encoding isozyme LH3, DNA and amino acid sequence determination and analysis, quantitative expression analysis
A5A6S1;, A5A6S2;, A5A6S3;
gene Plod1, quantitative RT-PCR enzyme expression analysis; gene Plod3, quantitative RT-PCR enzyme expression analysis; genes Plod2a and Plod2b, quantitative RT-PCR enzyme expression analysis
gene Plod1, real-time RT-PCR enzyme expression analysis; gene Plod2, real-time RT-PCR enzyme expression analysis; gene Plod3, real-time RT-PCR enzyme expression analysis
gene PLOD2
gene PLOD2, binding of the TGFbeta1 pathway related transcription factors SMAD3 and SP1-mediated TGFbeta1 enhanced PLOD2 expression and might be correlated to an increase of acetylated histone H3 and H4 at the PLOD2 promoter. Depletion of SMAD3 reduces gene PLOD2 acetylated H3 and H4, histone methylation marks at the PLOD2 promoter depicted an increase of the active histone mark H3K79me2, a decrease of the repressive H4K20me3 mark, but no role for the generally strong transcription-related modifications: H3K4me3, H3K9me3 and H3K27me3. TGFbeta1 induces a SP1- and SMAD3-dependent recruitment of histone modifying enzymes to the PLOD2 promoter other than the currently known TGFbeta1 downstream co-activators and epigenetic modifications. Quantitative real-time PCR enzyme expression analysis
-
gene PLOD2, DNA and amino acid sequence determination and analysis
gene PLOD2, encoding for splice variants LH2a and LH2b, quantitative expression analysis of LH2b
-
gene PLOD2, large-scale transfection with N-terminal His8 and human growth hormone (hGH) tags transfection with polyethylenimine and recombinant expression of wild-type enzyme, N-terminally truncated enzyme mutant, and of inactive mutant D689A in engineered CHO cells and secretion to the cell culture medium
-
gene PLOD3
-
gene PLOD3, DNA and amino acid sequence determination and analysis, quantitative expression analysis
gene PLOD3, expression in COS-7 and HT-1080 cells, the enzyme is secreted into the culture medium, expression of enzyme mutants in COS-7 cells
gene Plod3, expression of recombinant V5/His-tagged LH3 protein in HEK-293 cells
genes PLOD1 and PLOD2, real-time quantitative reverse transcription PCR
-
genomic structure of isozyme LH2, expression analysis of the 2 splicing variants of isozyme LH2
overexpression in HT-1080 cells, expression of fragments in Escherichia coli
quantitative LH gene expression analysis by realtime PCR, genes LH2a and LH2b DNA and amino acid sequence analysis using RT-PCR
-
transient transfection of HA-tagged LH1 in 714 mouse embryonic fibroblasts
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
cannabidiol stimulates mRNA expression of Plod1 in primary osteoblast cultures; DELTA9-tetrahydrocannabinol stimulates mRNA expression of Plod2 in primary osteoblast cultures
gene expressions of isozymes LH1, LH2b and LOXL2 are significantly upregulated by 1,25(OH)2D3, the active form of vitamin D
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hypoxia-inducible factor 1 activates transcription of the PLOD1 and PLOD2 genes in hypoxic breast cancer cells, knockdown of HIF-1a expression blocks PLOD1 and PLOD2 induction under hypoxic conditions
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in osteoarthritic human synovial fibroblasts, the enzyme expression is induced by TGF-beta via kinase ALK5, not ALK1, and Smad2/3P
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LH activity of LH3 is not reduced by mutations of this DXD motif
no induction of PLOD3 expression in osteoblasts by DELTA9-tetrahydrocannabinol and cannabidiol
overexpression of the TIA proteins in HEK-293 cells leads to an increase in levels of the LH2(long) spliced product in the minigene
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procollagen lysyl hydroxylase 2 expression is regulated by an alternative downstream transforming growth factor beta-1 activation mechanism. TGF-beta1 induces the enzyme, mechanism of TGFbeta1-enhanced PLOD2 expression, detailed overview
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reduction of TIA protein levels in mous embryonic fibroblasts and HEK-293 cells leads to a corresponding decrease in the LH2(long) spliced product in both the minigene and the endogenous gene
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D250A
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site-directed mutagenesis, the mutant shows unaltered lysyl hydroxylase activity, although reduced collagen glucosyltransferase activity, compared to the wild-type enzyme
D97A/D99A
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site-directed mutagenesis, the mutant cannot be expressed in Escherichia coli
H80A
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site-directed mutagenes, the mutant cannot be expressed in Escherichia coli
H825S/D827A
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site-directed mutagenesis, lysyl hydroxylase inactive mutant, that is still active as collagen glucosyltransferase
L78K
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site-directed mutagenesis, the mutant cannot be expressed in Escherichia coli
D250A
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site-directed mutagenesis, the mutant shows unaltered lysyl hydroxylase activity, although reduced collagen glucosyltransferase activity, compared to the wild-type enzyme
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D97A/D99A
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site-directed mutagenesis, the mutant cannot be expressed in Escherichia coli
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H80A
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site-directed mutagenes, the mutant cannot be expressed in Escherichia coli
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H825S/D827A
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site-directed mutagenesis, lysyl hydroxylase inactive mutant, that is still active as collagen glucosyltransferase
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L78K
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site-directed mutagenesis, the mutant cannot be expressed in Escherichia coli
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C144I
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isoform 3, reduces glycosyltransferase activity
L208I
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isoform 3, reduces glycosyltransferase activity
A1011G
naturally occuring heterozygous polymorphism in gene PLOD3
A195G
naturally occuring polymorphism in gene PLOD3
A434G
naturally occuring polymorphism in gene PLOD3
C882T
naturally occuring heterozygous polymorphism in gene PLOD3
D689A
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inactive mutant
E542A
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site-directed mutagenesis
E542A/E547A
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site-directed mutagenesis
E542A/E547A/E574A/E579A
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site-directed mutagenesis
E542A/E547A/E574A/E579A/E560A
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site-directed mutagenesis
E542A/H546L/E547A
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site-directed mutagenesis
E542A/Q543L/Y544F/E547A/E574A
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site-directed mutagenesis
E547A
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site-directed mutagenesis
E579A/E560A
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site-directed mutagenesis
G597V
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naturally occuring recessive point mutation, leads to abnormal folding and oligomerization of the mutant enzyme, which shows over 95% reduced activity compared to the wild-type enzyme
K541M
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site-directed mutagenesis
K541M/E542A
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site-directed mutagenesis
K694G
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site-directed mutagenesis, point mutation is introduced in the LH1 part of the expression construct comprising 40 amino acid residues of the C-terminal end of isozyme LH1, responsible for endoplasmic reticulum localization, fused to human cathepsin D and c-Myc-tagged, the exchange of the charged residue and deletion of 8 amino acids of the last 40 residues at the enzymes' C-terminal end has no effect on retention efficiency of the reporter protein, but deletion of the next 8 amino acid residues, leaving 24 residues, increases the secretion level of enzyme from the cell
N223S
site-directed mutagenesis, the mutant shows 50% reduced lysylhydroxylase activity, while the glycosyltransferase activity is almost abolished
Q543L/Y544F
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site-directed mutagenesis
R594H
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naturally occuring recessive point mutation, leads to abnormal folding and oligomerization of the mutant enzyme, which shows over 95% reduced activity compared to the wild-type enzyme
R693Q
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site-directed mutagenesis, point mutation is introduced in the LH1 part of the expression construct comprising 40 amino acid residues of the C-terminal end of isozyme LH1, responsible for endoplasmic reticulum localization, fused to human cathepsin D and c-Myc-tagged, the exchange of the charged residue and deletion of 8 amino acis of the last 40 residues at the enzymes' C-terminal end has no effect on retention efficiency of the reporter protein, but deletion of the next 8 amino acid residues, leaving 24 residues, increases the secretion level of enzyme from the cell
T604I
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naturally occuring recessive point mutation, leads 70-92% reduced activity, dependent on the 2-oxoglutarate concentration, compared top the wild-type due to a 10fold increase in the Km for 2-oxoglutarate, the mutant shows unaltered folding and oligomerization. The Km values of the T604I mutant for the peptide substrate, Fe2+, and ascorbate are identical to those of the wild-type
W446G
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naturally occurring mutation T1360G in a highly conserved region of exon 13 of isozyme LH1 in skin fibroblasts is predicted to lead to the W446G exchange in heterozygous Ehlers-Danlos syndrome type IVA, leads to loss of enzyme activity and causes the pathogenic effect probably due to incorect folding of isozyme LH1, structure-function analysis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
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PLOD expression is associated with human breast cancer progression, PLOD2 expression is specifically prognostic in breast cancer
medicine