Information on EC 1.5.5.2 - proline dehydrogenase

for references in articles please use BRENDA:EC1.5.5.2
Word Map on EC 1.5.5.2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY hide
1.5.5.2
-
RECOMMENDED NAME
GeneOntology No.
proline dehydrogenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-proline + a quinone = (S)-1-pyrroline-5-carboxylate + a quinol
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
(5R)-carbapenem carboxylate biosynthesis
-
-
L-citrulline biosynthesis
-
-
L-Ndelta-acetylornithine biosynthesis
-
-
L-proline degradation
-
-
proline to cytochrome bo oxidase electron transfer
-
-
(5R)-carbapenem carboxylate biosynthesis
-
-
proline metabolism
-
-
Arginine and proline metabolism
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
L-proline:quinone oxidoreductase
A flavoprotein (FAD). The electrons from L-proline are transferred to the FAD cofactor, and from there to a quinone acceptor [3]. In many organisms, ranging from bacteria to mammals, proline is oxidized to glutamate in a two-step process involving this enzyme and EC 1.2.1.88, L-glutamate gamma-semialdehyde dehydrogenase. Both activities are carried out by the same enzyme in enterobacteria.
CAS REGISTRY NUMBER
COMMENTARY hide
9050-70-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
collected from Yuanmou, Yunnan Province, China
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
Uniprot
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
mungbean
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
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
hydroxy-L-proline + acceptor + H2O
?
show the reaction diagram
-
3%5 the rate of L-proline
-
-
?
L-proline + 2,6-dichlorophenolindophenol + H2O
(S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
L-proline + 2,6-dichlorophenolindophenol + H2O
DELTA1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
L-proline + 2,6-dichlorphenol-indophenol
(S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorphenol-indophenol
show the reaction diagram
L-proline + a quinone
(S)-1-pyrroline-5-carboxylate + a quinol
show the reaction diagram
L-proline + acceptor
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
L-proline + acceptor + H2O
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
L-proline + cytochrome c
(S)-1-pyrroline-5-carboxylate + reduced cytochrome c
show the reaction diagram
L-proline + FAD + H2O
(S)-1-pyrroline-5-carboxylate + FADH2
show the reaction diagram
-
-
-
-
?
L-proline + ferricyanide + H2O
(S)-1-pyrroline-5-carboxylate + ferrocyanide
show the reaction diagram
-
-
-
?
L-proline + NAD+
(S)-1-pyrroline-5-carboxylate + NADH
show the reaction diagram
L-proline + NAD+ + H2O
(S)-1-pyrroline-5-carboxylate + NADH
show the reaction diagram
-
-
-
?
L-proline + oxidized dichlorophenolindophenol
1-pyrroline-5-carboxylate + reduced dichlorophenolindophenol
show the reaction diagram
-
-
-
?
L-proline + oxidized phenazine methosulfate
(S)-1-pyrroline-5-carboxylate + reduced phenazine methosulfate
show the reaction diagram
-
-
-
-
?
L-thiazolidine-4-carboxylate + acceptor + H2O
N-formylcysteine + reduced acceptor
show the reaction diagram
-
-
-
?
trans-4-hydroxy-L-proline + oxidized dichlorophenolindophenol
?
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
L-proline + a quinone
(S)-1-pyrroline-5-carboxylate + a quinol
show the reaction diagram
L-proline + acceptor
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
L-proline + acceptor + H2O
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
L-proline + oxidized phenazine methosulfate
(S)-1-pyrroline-5-carboxylate + reduced phenazine methosulfate
show the reaction diagram
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATP
O59088 and O59089
-
coenzyme Q1
phenazine methosulfate
-
-
[4Fe-4S]-center
O59088 and O59089
-
additional information
-
the recombinant holo form of MBP-tagged TtProDH, as produced in Escherichia coli TOP10 cells, contains about three times more FMN than FAD. The crystal structure of TtProDH variant DELTAABC, which lacks helices alphaA, alphaB and alphaC, shows no electron density for an AMP moiety of the cofactor. ProDH adopts a distorted (betalpha)8 TIM-barrel fold and is the only known TIM-barrel enzyme that contains an FAD cofactor. One cofactor molecule per enzyme subunit
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
the alpha-subunit contains the [2Fe-2S] iron-sulfur flavoprotein. The gamma subunit is a typical [8Fe-8S] ferredoxin
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-Thenoyltrifluoroacetone
5-ethylpentyl-barbituric acid
-
membrane-associated enzyme, not soluble enzyme
acetate
competitive
antimycin A
cyanide
-
only membrane-associated enzyme, not soluble enzyme
D-lactate
GuHCl
-
enzyme unfolding at 1 M, 0.5 M is not enough for proper unfolding. The fusion protein forms visible aggregates due to unfolding of MBP
-
L-Azetidine-2-carboxylate
-
competitive
L-lactate
L-proline
L-tetrahydro-2-furoic acid
L-thiazolidine-4-carboxylate
-
competitive
Lactate
-
-
N-propargylglycine
-
irreversibly inactivates PutA by covalently linking the flavin N(5) atom to the epsilon-amino of Lys329. Inactivation locks PutA into a conformation that may mimic the proline-reduced, membrane-associated form
pyrrolidone-5-carboxylate
-
pyruvate
-
-
rotenone
succinate
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0034
2,6-dichlorophenolindophenol
-
wild type enzyme, at 25°C and pH 7.1 in 20 mM Tris-HCl
0.01658 - 340
L-proline
0.0235 - 42
NAD+
190 - 750
trans-4-hydroxy-L-proline
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5 - 100.6
L-proline
2 - 2.9
trans-4-hydroxy-L-proline
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
21.9 - 79.5
2,6-dichlorophenolindophenol
1.438 - 90.32
coenzyme Q1
0.125
L-hydroxyproline
-
in 200 mM Tris-HCl buffer (pH 8.0), at 50°C
0.00021 - 198
L-proline
31 - 235
NAD+
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.1
D-lactate
-
-
20
L-Azetidine-2-carboxylate
-
-
1 - 1.4
L-lactate
0.084 - 7.9
L-tetrahydro-2-furoic acid
3.3
pyruvate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.014
O59088 and O59089
pH and temperature not specified in the publication
0.68
bifunctional dye-linked L-proline/NADH dehydrogenase complex
0.84
-
purified recombinant apo-enzyme, pH 7.4, 25°C, with 2,6-dichlorphenol-indophenol
1.24
-
25°C, pH 7.5, proline oxidase activity of the bifunctional proline dehydrogenase/L-glutamate gamma-semialdehyde dehydrogenase
4
-
purified recombinant MBP-tagged enzyme, pH 8.0, 25°C
4.74
-
purified recombinant holo-enzyme, pH 7.4, 25°C, with 2,6-dichlorphenol-indophenol
4.83
-
purified recombinant apo-enzyme reconstituted with FAD, pH 7.4, 25°C, with 2,6-dichlorphenol-indophenol
4.95
-
purified recombinant apo-enzyme reconstituted with FMN, pH 7.4, 25°C, with 2,6-dichlorphenol-indophenol
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 9
enzyme ProDH shows a good activity in the range of pH 7.0-9.0, and optimum pH at pH 8.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 70
recombinant ProDH, highest activity at 30°C, loss of activity above 30°C, near inactivation at 70°C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.27
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
high enzyme expression level
Manually annotated by BRENDA team
low enzyme expression level
Manually annotated by BRENDA team
low enzyme expression level
Manually annotated by BRENDA team
high enzyme expression level
Manually annotated by BRENDA team
low enzyme expression level
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
transcriptional induction of the enzyme causes changes in expression levels of other mitochondrial enzymes. Activity of glutamate dehydrogenase is substantially increased, and activity of D-lactate dehydrogenase is increased
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8000
alphabetagammadelta, 1 * 54000 + 1 * 43000 + 1 * 19000 + 1 * 8000, the beta-subunit catalyzes the dye-linked L-proline dehydrogenase reaction by itself, the alpha-subunit exhibts dye-linked NADH dehydrogenase activity, SDS-PAGE
19000
alphabetagammadelta, 1 * 54000 + 1 * 43000 + 1 * 19000 + 1 * 8000, the beta-subunit catalyzes the dye-linked L-proline dehydrogenase reaction by itself, the alpha-subunit exhibts dye-linked NADH dehydrogenase activity, SDS-PAGE
37000
-
2 * 37000, SDS-PAGE
40000
-
alpha,beta, 2 * 50000 + 2 * 40000, SDS-PAGE
42700
O59088 and O59089
-
43000
alphabetagammadelta, 1 * 54000 + 1 * 43000 + 1 * 19000 + 1 * 8000, the beta-subunit catalyzes the dye-linked L-proline dehydrogenase reaction by itself, the alpha-subunit exhibts dye-linked NADH dehydrogenase activity, SDS-PAGE
50000
-
alpha,beta, 2 * 50000 + 2 * 40000, SDS-PAGE
54000
alphabetagammadelta, 1 * 54000 + 1 * 43000 + 1 * 19000 + 1 * 8000, the beta-subunit catalyzes the dye-linked L-proline dehydrogenase reaction by itself, the alpha-subunit exhibts dye-linked NADH dehydrogenase activity, SDS-PAGE
55300
O59088 and O59089
-
76000
gel electrophoresis
78675
-
recombinant MBP-tagged enzyme, sequence calculation
78677
-
recombinant MBP-tagged enzyme, mass spectrometry
120000
bifunctional dye-linked L-proline/NADH dehydrogenase complex, gel filtration
137000
-
2 * 137000, bifunctional enzyme: proline dehydrogenase/L-glutamate gamma-semialdehyde dehydrogenase, SDS-PAGE
260000
-
gel filtration
293000
-
gel filtration, bifunctional enzyme: proline dehydrogenase/L-glutamate gamma-semialdehyde dehydrogenase, sucrose density centrifugation
440000
O59088 and O59089
gel filtration
additional information
-
the MBP tag inhibits the self-association of TtProDH, preventing to a large extent the formation of insoluble protein aggregates
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterooctamer
O59088 and O59089
4 * 55300 + 4 * 42700, calculated from amino acid sequence
homodimer
-
2 * 37000, SDS-PAGE
octamer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2.15 A resolution structure of the PRODH inactivated by N-propargylglycine. The initial events involved in broadcasting the reduced flavin state to the distal membrane-binding domain include major reorganization of the flavin ribityl chain, severe butterfly bending of the isoalloxazine ring, and disruption of an electrostatic network involving the flavin N(5) atom, Arg431, and Asp370. The active site is incompletely assembled in the absence of the substrate, and the binding of proline draws together conserved residues in helix 8 and the beta1-alphal loop to complete the active site
-
hanging drop method
mutant Y540S
-
sitting-drop vaor diffusion method, structures of the PutA PRODH domain complexed with competitive inhibitors, acetate, L-lactate and L-tetrahydro-2-furoic acid
purified TtProDH mutant variant DELTAABC lacking helices alphaA, alphaB and alphaC, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 10
-
50°C, 10 min, stable
392570
4.5 - 9
stable
658668
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
50% loss of activity of wild-type PutA86-669 after 1 h, 50% loss of of L432P mutant form of PutA86-669 after 8 min
80
-
120 min, 40% of activity remains
additional information
-
enzyme unfolding experiments using recombinant MBP-tagged enzyme in 20 mM Hepes, pH 8.0, at 60-95°C, the MBP-tagged enzyme unfolds at lower temperature around 56°C due to unfolding of the MBP tag
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stabilized by 1 M NaCl, KCl
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isolated mitochondria
nickel affinity column chromatography
-
partially by isolation of mitochondria
recombinant active His6-tagged enzyme from Escherichia coli strain BL21-CodonPlus (DE3) by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and dialysis to homogeneity
recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and dialysis to homogeneity, the PH1751 protein tends to aggregate during dialysis; recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and dialysis to homogeneity, the PH1751 protein tends to aggregate during dialysis
recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and dialysis to homogeneity; recombinant His-tagged enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIPL by nickel affinity chromatography and dialysis to homogeneity
recombinant MBP-tagged enzyme from Escherichia coli, gel filtration, and tag cleavage by trypsin, followed by ultrafiltration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene JcProDH, DNA and amino acid sequence determination and analysis, sequence analysis, phylogenetic analysis, reverse transcription PCR expression analysis, recombinant functional expression in Saccharomyces cerevisiae strain INVSc1
gene POX, stable recombinant expression in DLD-1 colorectal cancer cells
gene ProDH, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)plysS
gene product PutA functions as proline dehydrogenase and as repressor of genes putP and putA
-
gene Tc00.1047053506411.30, sequence comparisons, recombinant expression of the active His6-tagged enzyme in Escherichia coli strain BL21-CodonPlus (DE3) cells, reverse transcription PCR (RT-PCR) and quantitative real-time PCR (qRT-PCR) expression analysis, functional complementation of ProDH-deficient, PUT1 mutant Saccharomyces cerevisiae strain YLR142W. TcPRODH gene expression in the mutant diminishes free intracellular proline levels and an enhances sensitivity to oxidative stress
gene TT_C1214, recombinant expression of functional and soluble MBP-tagged enzyme in Escherichia coli TOP10 cells. The MBP tag inhibits the self-association of TtProDH, preventing to a large extent the formation of insoluble protein aggregates
-
recombinant expression of the holo form of MBP-tagged TtProDH in Escherichia coli TOP10 cells, the recombinant enzyme contains about three times more FMN than FAD. The apoenzyme is produced by riboflavin auxotrophic Escherichia coli strain BSV11 and can be successfully reconstituted with FAD or FMN. The recombinant apo-enzyme reconstituted with FAD or FMN shows equal specific activities as holo-enzyme
-
sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3)-RIPL
sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3)-RIPL; sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-CodonPlus(DE3)-RIPL
wild-type PutA86-601 and site-directed mutants
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
isoform ProDH1 expression is dependent on salicylic acid (stimulation at 0.05-2.5 mM) and an increase in enzyme activity is detected in cells destined to die
-
L-proline treatment induces ProDH in planta. Induction of D-lactate dehydrogenase activity allows rapid upregulation of ProDH activity during the short-term stress response in plants. Transcriptional induction of the enzyme causes changes in expression levels of other mitochondrial enzymes
-
ProDH activity is high for genotypes in which ProDH, most likely ProDH1, is strongly induced by L-proline
proline and NaCl induce gene expression
-
salicylic acid has no effect on the isoform ProDH2 mRNA level, even at 2.5 mM
-
sucrose represses ProDH2 expression
-
TcPRODH mRNA and protein expression are strongly upregulated in the intracellular epimastigote, a stage which requires an external supply of proline
the expression of JcProDH increases in leaves experiencing environmental stress such as cold (5°C), heat (42°C), salt (300 mM), and drought (30% PEG 6000)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D370A
-
mutant form of PutA86-601. The ratio of turnover-number to Km-value is 2.8fold lower than the ratio of the wild-type PutA86-601
D370A/Y540F
-
mutant form of PutA86-601. The ratio of turnover-number to Km-value is 15fold lower than the ratio of the wild-type PutA86-601
L432P
mutant form of PutA86-669, 5fold decrease in turnover-number and a severe loss in thermostability
W194F
-
mutation in truncated enzyme containing residues 86-601, ratio of kcat to Km-value drops by 50%
W194F/W211F
-
mutation in truncated enzyme containing residues 86-601, ratio of kcat to Km-value drops by 95%
W211F
-
mutation in truncated enzyme containing residues 86-601, ratio of kcat to Km-value drops by 80%
Y540A
-
a construct consisting of residue 86-630 of PutA is used
Y540F
-
mutant form of PutA86-601. The ratio of turnover-number to Km-value is 5.8fold lower than the ratio of the wild-type PutA86-601
Y540S
-
a construct consisting of residue 86-630 of PutA is used
Y203F
-
the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme
F10E/L12E
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (10211 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.