Any feedback?
Information on EC 1.5.1.2 - pyrroline-5-carboxylate reductase and Organism(s) Homo sapiens and UniProt Accession P32322
for references in articles please use BRENDA:EC1.5.1.2Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
1.5.1.2 pyrroline-5-carboxylate reductaseIUBMB Comments
Also reduces 1-pyrroline-3-hydroxy-5-carboxylate to L-hydroxyproline.
Specify your search results
Word Map
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
Synonyms
pycr1, proline oxidase, pyrroline-5-carboxylate reductase, p5cr, p5c reductase, 1-pyrroline-5-carboxylate reductase, delta1-pyrroline-5-carboxylate reductase, l-proline oxidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
1-pyrroline-5-carboxylate reductase
-
-
-
-
L-proline oxidase
-
-
-
-
L-proline-NAD(P)+ 5-oxidoreductase
-
-
-
-
L-proline:NAD(P)+ 5-oxidoreductase
-
-
-
-
NADPH-L-DELTA'-pyrroline carboxylic acid reductase
-
-
-
-
P5C reductase
P5CR
proline oxidase
pyrroline-5-carboxylate reductase
reductase, pyrroline-5-carboxylate
-
-
-
-
P5C reductase
-
-
-
-
P5CR
-
-
-
-
proline oxidase
-
-
-
-
pyrroline-5-carboxylate reductase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-proline + NAD(P)+ = 1-pyrroline-5-carboxylate + NAD(P)H + H+
enzyme PYCR2 kinetics suggest a sequential binding mechanism with L-P5C binding before NAD(P)H and NAD(P)+ releasing before L-Pro
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
oxidation
reduction
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
L-proline:NAD(P)+ 5-oxidoreductase
Also reduces 1-pyrroline-3-hydroxy-5-carboxylate to L-hydroxyproline.
CAS REGISTRY NUMBER
COMMENTARY
9029-17-8
-
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
L-pipecolate + NADP+
DELTA1-piperideine 6-carboxylate + NADPH + H+
-
-
-
?
L-Pro + NAD(P)+
1-pyrroline-5-carboxylate + NAD(P)H
-
the enzyme plays a role as a downstream effector in p53-mediated apoptosis of renal carcinoma cells
-
-
?
L-proline + NAD(P)+
1-pyrroline-5-carboxylate + NAD(P)H + H+
-
-
-
-
r
piperideine-6-carboxylate + NADH + H+
L-pipecolic acid + NAD+
-
-
L-pipecolic acid is the sole product
-
ir
piperideine-6-carboxylate + NADPH + H+
L-pipecolic acid + NADP+
-
-
L-pipecolic acid is the sole product
-
ir
pyrroline-5-carboxylate + NAD(P)H + H+
L-proline + NAD(P)+
-
third enzyme in proline pathway
-
-
?
pyrroline-5-carboxylate + NAD(P)H + H+
L-proline + NAD(P)+
-
linkage to hexose monophosphate pathway
-
-
?
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
L-pipecolate + NADP+
DELTA1-piperideine 6-carboxylate + NADPH + H+
-
-
-
?
L-Pro + NAD(P)+
1-pyrroline-5-carboxylate + NAD(P)H
-
the enzyme plays a role as a downstream effector in p53-mediated apoptosis of renal carcinoma cells
-
-
?
L-proline + NAD(P)+
1-pyrroline-5-carboxylate + NAD(P)H + H+
-
-
-
-
r
pyrroline-5-carboxylate + NAD(P)H + H+
L-proline + NAD(P)+
-
third enzyme in proline pathway
-
-
?
pyrroline-5-carboxylate + NAD(P)H + H+
L-proline + NAD(P)+
-
linkage to hexose monophosphate pathway
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
PYCR2 does not show a strong preference for NADH or NADPH
-
additional information
-
PYCR2 does not show a strong preference for NADH or NADPH
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
([[(3,5-dibromophenyl)methyl]amino]methylene)bis(phosphonic acid)
competitive inhibition against both 1-pyrroline 5-carboxylate and NADPH. Growth of myelogenous erythroleukemic K562 and epithelial breast cancer MDA-MB-231 cell lines is progressively impaired by concentrations ranging from 1-100 microM
-
([[(3,5-dichlorophenyl)methyl]amino]methylene)bis(phosphonic acid)
competitive inhibition against both 1-pyrroline 5-carboxylate and NADPH
-
N-formyl-L-proline
competitive. Compound phenocopies the PYCR1 knockdown in MCF10A H-RASV12 breast cancer cells by inhibiting de novo proline biosynthesis and impairing spheroidal growth
-
stearoyl-CoA
-
competitive inhibitor of P5CR NAD(P)H-dependent thioproline dehydrogenase activity
L-proline
at 0-500 microM, the value of KMapp increases whereas Vlim remains fairly unchanged, which is consistent with competitive inhibition
proline
-
competitive inhibitor of P5CR NAD(P)H-dependent thioproline dehydrogenase activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.667
1-pyrroline-5-carboxylate
cosubstrate NADPH, wild-type, pH 7.5, 37°C
0.317
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADH
0.32
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADH, pH 7.5, 37°C
0.33
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADPH, pH 7.5, 37°C
0.334
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADPH
0.949
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADPH
0.99
1-pyrroline-5-carboxylate
wild-type, cosubstrate NADPH, pH 7.5, 37°C
1.315
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADH
1.32
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADH, pH 7.5, 37°C
1.499
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADPH
1.5
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADPH, pH 7.5, 37°C
1.509
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADH
0.151
NAD+
-
wild type enzyme, 1 mM fixed thioproline, thioproline dehydrogenase activity assay
0.235
NAD+
-
E221A mutant, 1 mM fixed thioproline, thioproline dehydrogenase activity assay
0.034
NADH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
0.298
NADH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
0.953
NADH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
0.48
NADP+
-
E221A mutant, 1 mM fixed thioproline, thioproline dehydrogenase activity assay
3.06
NADP+
-
wild type enzyme, 1 mM fixed thioproline, thioproline dehydrogenase activity assay
0.119
NADPH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
0.216
NADPH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
0.537
NADPH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
additional information
additional information
steady-state kinetics of the wild-type enzyme and mutants
-
additional information
additional information
-
steady-state kinetics of the wild-type enzyme and mutants
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.08
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADH, pH 7.5, 37°C
0.08
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADH
0.13
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADPH, pH 7.5, 37°C
0.13
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADPH
3.1
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADPH, pH 7.5, 37°C
3.1
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADPH
3.2
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADH, pH 7.5, 37°C
3.2
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADH
26
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADPH
61.3
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADH
0.015
NADH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
5.9
NADH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
47.9
NADH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
1.5
NADPH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
1.8
NADPH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
24
NADPH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.25
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADH, pH 7.5, 37°C
0.25
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADH
0.39
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R119C, with NADPH
0.4
1-pyrroline-5-carboxylate
mutant R119C, cosubstrate NADPH, pH 7.5, 37°C
2.07
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADPH
2.1
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADPH, pH 7.5, 37°C
2.4
1-pyrroline-5-carboxylate
mutant R251C, cosubstrate NADH, pH 7.5, 37°C
2.43
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant mutant R251C, with NADH
27.4
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADPH
40.62
1-pyrroline-5-carboxylate
pH 7.5, 37°C, recombinant wild-type enzyme, with NADH
0.441
NADH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
6.19
NADH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
160.74
NADH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
2.793
NADPH
pH 7.5, 37°C, recombinant mutant R119C, with 1-pyrroline-5-carboxylate
15.13
NADPH
pH 7.5, 37°C, recombinant mutant R251C, with 1-pyrroline-5-carboxylate
111.11
NADPH
pH 7.5, 37°C, recombinant wild-type enzyme, with 1-pyrroline-5-carboxylate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.145
L-proline
pH 7.5, 37°C, recombinant wild-type enzyme, competitive inhibition
additional information
additional information
product(s) inhibition kinetics are analyzed by global fitting of the data to different inhibition models
-
additional information
additional information
-
product(s) inhibition kinetics are analyzed by global fitting of the data to different inhibition models
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.19
cyclopentanecarboxylate
Homo sapiens
pH 7.8, temperature not specified in the publication
2.25
L-tetrahydro-2-furoic acid
Homo sapiens
pH 7.8, temperature not specified in the publication
0.44
L-thiazolidine-2-carboxylate
Homo sapiens
pH 7.8, temperature not specified in the publication
-
0.6
L-thiazolidine-4-carboxylate
Homo sapiens
pH 7.8, temperature not specified in the publication
0.1
N-formyl-L-proline
Homo sapiens
pH 7.8, temperature not specified in the publication
-
0.00039
([[(3,5-dibromophenyl)methyl]amino]methylene)bis(phosphonic acid)
Homo sapiens
substrate NADPH, pH 7.5, 30°C
-
0.00048
([[(3,5-dibromophenyl)methyl]amino]methylene)bis(phosphonic acid)
Homo sapiens
substrate NADH, pH 7.5, 30°C
-
0.00074
([[(3,5-dichlorophenyl)methyl]amino]methylene)bis(phosphonic acid)
Homo sapiens
substrate NADPH, pH 7.5, 30°C
-
0.00078
([[(3,5-dichlorophenyl)methyl]amino]methylene)bis(phosphonic acid)
Homo sapiens
substrate NADH, pH 7.5, 30°C
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
14
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
full-length sequence is amplified from a hepatoma cell line cDNA library
-
colorectal cancer cell, DLD-1 Tet-Off POX cells are generated for controlled expression of proline oxidase
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
enzymes that reduce DELTA1-pyrroline-5-carboxylate and DELTA1-piperideine-6-carboxylate are aldimine reductases whereas enzymes that reduce DELTA1-piperideine-2-carboxylate and DELTA1-pyrroline-2-carboxylate (P2C/Pyr2C) are ketimine reductases (KRs)
malfunction
homozygous mutations in human PYCR2 lead to postnatal microcephaly and hypomyelination, including hypomyelinating leukodystrophy type 10. PYCR2 mutations are detected in patients with lethal microcephaly, loss of PYCR2 is proposed to impair mitochondria function and disrupt oxidative balance, namely NAD(P)+ levels. The R251C variant displays the least overall regular alpha-helical character of the PYCR2 proteins, yet the R251C variant also displays strong overall regular beta-strand or beta-sheet character
metabolism
physiological function
metabolism
sequential binding mechanism with 1-pyrroline-5-carboxylate binding before NAD(P)H and NAD(P)+ releasing before L-Pro
metabolism
mammalian tissues contain a Pyr5C/P6C reductase that is distinct from Pyr2C/P2C reductase. In higher eukaryotes Pyr5C is also generated by a mitochondrial isozyme Pyr5C synthetase (isozyme PYCR1, UniProt ID P32322). Lysine/ornithine catabolism and interconnected pathways with 5-carbon amino acid (L-glutamate gamma-semialdehyde, L-glutamate, L-ornithine, L-proline) metabolism in mammalian tissues, overview. Although the ketimine reductase (KR) is important in the formation of L-pipecolate in the brain, it is also an important source of L-proline. This proline (via proline oxidase) in turn is an important source of Pyr5C and hence of glutamate and to a lesser extent ornithine
metabolism
the enzyme is involved in the proline metabolism in the mitochondrion, overview
physiological function
-
treatment with oxidized low-density lipoproteins decreases the cell number per field and causes the cells to round up. Knock-down of proline oxidase via small interfering RNA further reduces viability of cancer cells treated with oxidized low-density lipoproteins, decreases oxidized low-density lipoproteins-associated reactive oxygen species generation, decreases autophagy measured via beclin-1 protein level and light-chain 3 protein-I into LC3-II conversion. Single proline oxidase overexpression is sufficient to activate autophagy. It leads to autophagosomes accumulation and increases conversion of LC3-I into LC3-II.Beclin-1 gene expression is directly dependent on proline oxidase catalytic activity, namely the generation of prloine-oxidase-dependent superoxide
physiological function
PYCR2 knockdown reduces cell viability, proliferation, migration, and invasion and increases apoptosis. PYCR2 knockdown increases Bax, cleaved caspase-3, and cleaved PARP levels and decreases Bcl-2, MMP-2, MMP-9, p-PI3K, p-AKT, and p-mTOR levels in colorectal cancer cells
physiological function
L-proline has important roles in mammalian bioenergetics, cellular redox control, apoptosis and cancer. In mammals, Pyr5C reductase exists in two isoforms: a short form is present in the intestine that is feedback inhibited by ornithine and may be important in the synthesis of L-arginine from L-glutamate in the intestine. On the other hand, most other tissues express a long form of DELTA1-pyrroline-5-carboxylate reductase, which is thought to be important in the synthesis of L-proline from L-glutamate
physiological function
pyrroline-5-carboxylate reductase (PYCR in humans) catalyzes the final step of L-proline biosynthesis by catalyzing the reduction of L-DELTA1-pyrroline-5-carboxylate (L-P5C) to L-proline using NAD(P)H as the hydride donor. In humans, three isoforms PYCR1c, and PYCR3 are known. Isozyme PYCR3 is a cytosolic enzyme whereas PYCR1 and PYCR2 are identified as mitochondrial isozymes. PYCR2 not only drives cellular supply of L-proline but it is also important for redox cycling of NAD(P)H/NAD(P)+
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). P5CR1_HUMAN
319
0
33361
Swiss-Prot
Secretory Pathway (Reliability: 3)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
enzyme forms a concentration-dependent decamer in solution, sedimentation velocity data
additional information
-
enzyme forms a concentration-dependent decamer in solution, sedimentation velocity data
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
binary complexes of PYCR1 with NADPH or proline, at 1.9 A resolution, and a ternary complex containing NADPH and a P5C/proline analog, to 1.85 A resolution. NADPH is bound to the Rossmann fold. Structures provide a model of the Michaelis complex formed during hydride transfer
PYCR1 complexed with N-formyl-L-proline, inhibitor binding is accompanied by conformational changes in the active site, including the translation of an alpha-helix by 1 A
the protein is crystallized by the hanging-drop vapor-diffusion method at 37°C and diffraction data are obtained to a resolution of 2.8 A
2.8 A resolution structure of the pyrroline-5-carboxylate reductase apo enzyme, and its ternary complex with NADPH and substrate analog at 3.1 A
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
T238A
mutation in conserved residue, about 10fold decrease in catalytic efficiency
R119C
R251C
additional information
R119C
mutation identified in patient with microcephaly and hypomyelination, mutant is catalytically impaired
R119C
a naturally occuring mutation in patients with microcephaly and hypomyelination. The mutant is catalytically impaired, depending on whether NADPH or NADH is used, the catalytic efficiency of the R119C protein variant is 40 or 366 times lower than that of the wild-type enzyme
R251C
mutation identified in patient with microcephaly and hypomyelination, mutant is catalytically impaired and has a pronounced folding defect
R251C
a naturally occuring mutation in patients with microcephaly and hypomyelination. The mutant is catalytically impaired, depending on whether NADPH or NADH is used, the catalytic efficiency of the R119C protein variant is 7 or 26 times lower than that of the wild-type enzyme. The R251C protein variant has a pronounced folding defect. The R251C variant displays the least overall regular alpha-helical character of the PYCR2 proteins, yet the R251C variant also displays strong overall regular beta-strand or beta-sheet character
additional information
identification of a single homozygous region near the telomere of chromosome 17 in a cohort of patients with cutis laxa type 2. The single nucleotide change leads to a missense mutation adjacent to a slice junction in the gene encoding pyrroline-5-carboxylate reductase 1 which results in exon skipping and leads to deletion of reductase functional domain-coding region and an obligatory downstream frameshift
additional information
-
identification of a single homozygous region near the telomere of chromosome 17 in a cohort of patients with cutis laxa type 2. The single nucleotide change leads to a missense mutation adjacent to a slice junction in the gene encoding pyrroline-5-carboxylate reductase 1 which results in exon skipping and leads to deletion of reductase functional domain-coding region and an obligatory downstream frameshift
additional information
-
N-truncation results in an insoluble protein, C-truncation does not alter the activity
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
54 - 79
PYCR2 wild-type and mutant R119C exhibit fairly sharp unfolding transitions and similar Tm values of 69°C and 67°C, respectively, whereas mutant R251C exhibits a dramatically lower Tm value of 54°C. Incubation of each PYCR2 enzyme with product ligands, L-Pro and NAD+, individually or together has no effect on the observed Tm values
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography, dialysis, and ultrafiltration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
into a pET28a+ vector coding for a N-terminal His-tag for expression in Escherichia coli BL21DE3
a proline oxidase antisense vector is constructed by amplifying a part of the proline oxidase cDNA and cloning it in the mammalian expression vector pCI
-
cloned into the pET28a+ vector with a His6 tag at the N-terminus for expression in the Escherichia coli strain B834DE3
-
gene PYR2, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) pLysS
proline oxidase cDNA is cloned into the pAdtrack vector, it is used to generate a recombinant adenovirus for transfection
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
PI3K inhibition significantly reduces PYCR1 mRNA and protein expression. Nutrient stress induced by glucose deprivation also regulates PYCR1 expression
in a mass spectrometry-based screening of metabolites in tissue sections from 256 esophageal squamous cell carcinoma (ESCC) patients, a significant upregulation of L-proline metabolism is found in the cancer region compared to the normal epithelium and muscle regions. Immunohistochemistry staining of the ESCC tissue sections identifies PYCR2 as the most enriched metabolic enzyme in the cancer region, consistent with increased L-proline found in the same tissue region
oxidized low-density lipoproteins upregulate proline oxidase through nuclear receptor peroxisome proliferator-activated receptor gamma
-
upregulated micro-RNA miR-23b* in renal cancer is an important regulator of POX. Ectopic overexpression of miR-23b* in normal renal cells results in striking downregulation of POX, whereas POX expression increases markedly when endogenous miR-23b* is knocked down by its antagomirs in renal cancer cells
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
medicine
identification of a single homozygous region near the telomere of chromosome 17 in a cohort of patients with cutis laxa type 2. The single nucleotide change leads to a missense mutation adjacent to a slice junction in the gene encoding pyrroline-5-carboxylate reductase 1 which results in exon skipping and leads to deletion of reductase functional domain-coding region and an obligatory downstream frameshift
medicine
both PYCR1 mRNA and protein expression are significantly associated with tumor size, grade and invasive molecular subtypes of breast cancers. Higher PYCR1 mRNA levels are significantly associated with poor survival of breast cancer patients, regardless of estrogen receptor status. Inhibition of PYCR1 by small-hairpin RNA significantly reduces the growth and invasion capabilities of the cells, while enhancing the cytotoxicity of doxorubicin in breast cancer cell lines MCF-7 (estrogen receptor positive) and MDA-MB-231 (estrogen receptor negative). Chemotherapy significantly improves survival in early-stage breast cancer patients with low PYCR1 expression levels
medicine
PYCR1 expression is upregulated in different gastric cancer cohorts. High PYCR1 protein expression is correlated with advanced tumor stage, aggressive histological type and high Ki-67 index. High PYCR1 expression in gastric cancer tissues is an indicator of poor outcome in gastric cancer patients. In vitro, PYCR1 knockdown markedly attenuates gastric cancer cells growth and promoted apoptosis, while overexpression produces the opposite effects. PI3K/Akt axis is strongly correlated with PYCR1 expression and PIK3CB and AKT1 mRNA expression are positively associated with PYCR1 in gastric cancer tissues. PI3K inhibition significantly reduces PYCR1 mRNA and protein expression. Nutrient stress induced by glucose deprivation also regulates PYCR1 expression
medicine
-
overexpression of proline oxidase causes apoptotic cell death in a variety of cancer cells
medicine
-
proline oxidase can mediate apoptosis through generation of reactive oxygen species
medicine
-
micro-RNA miR-23b*, by targeting POX, may function as an oncogene. Pairs of human renal carcinoma and normal tissues show a negative correlation between miR-23b* and POX protein expression, providing its clinical corroboration. Ectopic overexpression of miR-23b* in normal renal cells results in striking downregulation of POX, whereas POX expression increases markedly when endogenous miR-23b* is knocked down by its antagomirs in renal cancer cells
medicine
PYCR2 is highly expressed in colorectal cancer patients and high PYCR2 expression is associated with advanced stage, adenocarcinoma, nodal metastasis, and poor survival rate. PYCR2 knockdown reduces cell viability, proliferation, migration, and invasion and increases apoptosis. PYCR2 knockdown increases Bax, cleaved caspase-3, and cleaved PARP levels and decreases Bcl-2, MMP-2, MMP-9, p-PI3K, p-AKT, and p-mTOR levels in colorectal cancer cells
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Donald, S.P.; Sun, X.Y.; Hu, C.A.A.; Yu, J.; Mei, J.M.; Valle, D.; Phang, J.M.
Proline oxidase, encoded by p53-induced gene-6, catalyzes the generation of proline-dependent reactive oxygen species
Cancer Res.
61
1810-1815
2001
Homo sapiens
Yeh, G.C.; Phang, J.M.
The function of pyrroline-5-carboxylate reductase in human erythrocytes
Biochem. Biophys. Res. Commun.
94
450-457
1980
Homo sapiens
Valle, D.; Downing, S.J.; Phang, J.M.
Proline inhibition of pyrroline-5-carboxylate reductase: differences in enzymes obtained from animal and tissue culture sources
Biochem. Biophys. Res. Commun.
54
1418-1424
1973
Cricetulus griseus, Homo sapiens, Rattus norvegicus
Maxwell, S.A.; Rivera, A.
Proline oxidase induces apoptosis in tumor cells, and its expression is frequently absent or reduced in renal carcinomas
J. Biol. Chem.
278
9784-9789
2003
Homo sapiens
Liu, Y.; Borchert, G.L.; Donald, S.P.; Surazynski, A.; Hu, C.A.; Weydert, C.J.; Oberley, L.W.; Phang, J.M.
MnSOD inhibits proline oxidase-induced apoptosis in colorectal cancer cells
Carcinogenesis
26
1335-1342
2005
Homo sapiens
Rivera, A.; Maxwell, S.A.
The p53-induced gene-6 (proline oxidase) mediates apoptosis through a calcineurin-dependent pathway
J. Biol. Chem.
280
29346-29354
2005
Homo sapiens
Pandhare, J.; Cooper, S.K.; Phang, J.M.
Proline oxidase, a proapoptotic gene, is induced by troglitazone: evidence for both peroxisome proliferator-activated receptor gamma-dependent and -independent mechanisms
J. Biol. Chem.
281
2044-2052
2006
Homo sapiens
Meng, Z.; Lou, Z.; Liu, Z.; Li, M.; Zhao, X.; Bartlam, M.; Rao, Z.
Crystal structure of human pyrroline-5-carboxylate reductase
J. Mol. Biol.
359
1364-1377
2006
Homo sapiens
Liu, Y.; Borchert, G.L.; Surazynski, A.; Hu, C.A.; Phang, J.M.
Proline oxidase activates both intrinsic and extrinsic pathways for apoptosis: the role of ROS/superoxides, NFAT and MEK/ERK signaling
Oncogene
25
5640-5647
2006
Homo sapiens
Meng, Z.; Lou, Z.; Liu, Z.; Hui, D.; Bartlam, M.; Rao, Z.
Purification, characterization, and crystallization of human pyrroline-5-carboxylate reductase
Protein Expr. Purif.
49
83-87
2006
Homo sapiens (P32322), Homo sapiens
Phang, J.M.; Pandhare, J.; Zabirnyk, O.; Liu, Y.
PPARgamma and proline oxidase in cancer
PPAR Res.
2008
542694
2008
Homo sapiens
Guernsey, D.L.; Jiang, H.; Evans, S.C.; Ferguson, M.; Matsuoka, M.; Nightingale, M.; Rideout, A.L.; Provost, S.; Bedard, K.; Orr, A.; Dube, M.P.; Ludman, M.; Samuels, M.E.
Mutation in pyrroline-5-carboxylate reductase 1 gene in families with cutis laxa type 2
Am. J. Hum. Genet.
85
120-129
2009
Homo sapiens (P32322), Homo sapiens
Zabirnyk, O.; Liu, W.; Khalil, S.; Sharma, A.; Phang, J.M.
Oxidized low-density lipoproteins upregulate proline oxidase to initiate ROS-dependent autophagy
Carcinogenesis
31
446-454
2010
Homo sapiens
Liu, W.; Zabirnyk, O.; Wang, H.; Shiao, Y.H.; Nickerson, M.L.; Khalil, S.; Anderson, L.M.; Perantoni, A.O.; Phang, J.M.
miR-23b targets proline oxidase, a novel tumor suppressor protein in renal cancer
Oncogene
29
4914-4924
2010
Homo sapiens
Christensen, E.M.; Patel, S.M.; Korasick, D.A.; Campbell, A.C.; Krause, K.L.; Becker, D.F.; Tanner, J.J.
Resolving the cofactor-binding site in the proline biosynthetic enzyme human pyrroline-5-carboxylate reductase 1
J. Biol. Chem.
292
7233-7243
2017
Homo sapiens (P32322), Homo sapiens
Struys, E.A.; Jansen, E.E.; Salomons, G.S.
Human pyrroline-5-carboxylate reductase (PYCR1) acts on delta(1)-piperideine-6-carboxylate generating L-pipecolic acid
J. Inherit. Metab. Dis.
37
327-332
2014
Homo sapiens
Hallen, A.; Cooper, A.J.
Reciprocal control of thyroid binding and the pipecolate pathway in the brain
Neurochem. Res.
42
217-243
2017
Homo sapiens (Q96C36)
Xiao, S.; Li, S.; Yuan, Z.; Zhou, L.
Pyrroline-5-carboxylate reductase 1 (PYCR1) upregulation contributes to gastric cancer progression and indicates poor survival outcome
Ann. Transl. Med.
8
937
2020
Homo sapiens (P32322), Homo sapiens
Patel, S.; Seravalli, J.; Liang, X.; Tanner, J.; Becker, D.
Disease variants of human Delta1-pyrroline-5-carboxylate reductase 2 (PYCR2)
Arch. Biochem. Biophys.
703
108852
2021
Homo sapiens (Q96C36), Homo sapiens
Ding, J.; Kuo, M.L.; Su, L.; Xue, L.; Luh, F.; Zhang, H.; Wang, J.; Lin, T.G.; Zhang, K.; Chu, P.; Zheng, S.; Liu, X.; Yen, Y.
Human mitochondrial pyrroline-5-carboxylate reductase 1 promotes invasiveness and impacts survival in breast cancers
Carcinogenesis
38
519-531
2017
Homo sapiens (P32322), Homo sapiens
Yin, F.; Huang, X.; Xuan, Y.
Pyrroline-5-carboxylate reductase-2 promotes colorectal cancer progression via activating PI3K/AKT/mTOR pathway
Dis. Markers
2021
9950663
2021
Homo sapiens (Q96C36)
Christensen, E.M.; Bogner, A.N.; Vandekeere, A.; Tam, G.S.; Patel, S.M.; Becker, D.F.; Fendt, S.M.; Tanner, J.J.
In crystallo screening for proline analog inhibitors of the proline cycle enzyme PYCR1
J. Biol. Chem.
295
18316-18327
2020
Homo sapiens (P32322), Homo sapiens
Forlani, G.; Sabbioni, G.; Ragno, D.; Petrollino, D.; Borgatti, M.
Phenyl-substituted aminomethylene-bisphosphonates inhibit human P5C reductase and show antiproliferative activity against proline-hyperproducing tumour cells
J. Enzyme Inhib. Med. Chem.
36
1248-1257
2021
Homo sapiens (P32322), Homo sapiens
EXTERNAL LINKS (specific for EC-Number 1.5.1.2)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
