Information on EC 1.6.1.2 - NAD(P)+ transhydrogenase (Re/Si-specific)

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

EC NUMBER
COMMENTARY hide
1.6.1.2
-
RECOMMENDED NAME
GeneOntology No.
NAD(P)+ transhydrogenase (Re/Si-specific)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
NADPH + NAD+ = NADP+ + NADH
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
NAD/NADH phosphorylation and dephosphorylation
-
-
Nicotinate and nicotinamide metabolism
-
-
NAD metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
NADPH:NAD+ oxidoreductase (Re/Si-specific)
The enzyme from heart mitochondria is Re-specific with respect to NAD+ and Si-specific with respect to NADP+ [cf. EC 1.6.1.1 NAD(P)+ transhydrogenase (Si-specific)].
CAS REGISTRY NUMBER
COMMENTARY hide
9014-18-0
not distinguished from EC 1.6.1.1
9072-60-0
not distinguished from EC 1.6.1.1
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene Nnt
-
-
Manually annotated by BRENDA team
formerly Rhodopseudomonas capsulata
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
the enzyme uses energy from the mitochondrial proton gradient to produce high concentrations of NADPH
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
H+/out + NADH + NADP+
H+/in + NAD+ + NADPH
show the reaction diagram
NADH + NADP+
NAD+ + NADPH
show the reaction diagram
NADH + NADP+
NADPH + NAD+
show the reaction diagram
NADH + oxidized 3-acetylpyridine adenine dinucleotide
NAD+ + reduced 3-acetylpyridine adenine dinucleotide
show the reaction diagram
-
-
-
-
?
NADH + thio-NADP+
NAD+ + thio-NADPH
show the reaction diagram
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
NADPH + 3-acetylpyridine adenine dinucleotide
NADP+ + reduced 3-acetylpyridine adenine dinucleotide
show the reaction diagram
-
-
-
-
?
NADPH + 3-acetylpyridine-NAD(P)+
NADP+ + 3-acetylpyridine-NAD(P)H
show the reaction diagram
-
-
-
-
r
NADPH + 3-acetylpyridine-NAD+
3-acetylpyridine-NADH + NADP+
show the reaction diagram
NADPH + NAD+
NADP+ + NADH
show the reaction diagram
NADPH + NAD+ + H+/in
NADP+ + NADH + H+/out
show the reaction diagram
-
-
-
r
NADPH + NAD+ + H+[side 1]
NADP+ + NADH + H+[side 2]
show the reaction diagram
-
-
-
-
?
NADPH + oxidized 3-acetylpyridine adenine dinucleotide
NADP+ + reduced 3-acetylpyridine adenine dinucleotide
show the reaction diagram
NMNH + thio-NADP+
NMN + thio-NADPH
show the reaction diagram
-
-
-
?
thio-NADH + NADP+
thio-NAD+ + NADPH
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
H+/out + NADH + NADP+
H+/in + NAD+ + NADPH
show the reaction diagram
NADH + NADP+
NAD+ + NADPH
show the reaction diagram
NADH + NADP+
NADPH + NAD+
show the reaction diagram
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
NADPH + NAD+
NADP+ + NADH
show the reaction diagram
NADPH + NAD+ + H+/in
NADP+ + NADH + H+/out
show the reaction diagram
Q24858
-
-
-
r
NADPH + NAD+ + H+[side 1]
NADP+ + NADH + H+[side 2]
show the reaction diagram
-
-
-
-
?
NADPH + oxidized 3-acetylpyridine adenine dinucleotide
NADP+ + reduced 3-acetylpyridine adenine dinucleotide
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD(P)+
NAD(P)H
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Li+
-
not clear whether this reflects a general salt effect or a Li+ specific effect
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2',5'-ADP
-
bacteriorhodopsin co-reconstituted enzyme, 36.8% inhibition of thio-NADP+ reduction by NADH in the dark, 34.4% in the light
2'-AMP
2,2'-dithiodipyridine
-
0.2 mM, 45% inhibition
2,2'-thiodiethanethiole
-
0.5 mM, 37% inhibition
2,4-Dinitrophenyl-3'-dephospho-CoA
-
competitive vs. NAD+, non-competitive vs. NADPH
2-(4-maleimidoanilino)-naphthalene-6-sulfonic acid
-
0.004 mM, 2 h incubation, 75% inhibition of reverse reaction catalyzed by A348C mutant enzyme, 95% inhibition of A390C mutant enzyme after 1 h, 90% inhibition of K424C mutant enzyme after 1 h, 55% inhibition of R425C mutant enzyme after 1h
-
3'-5'-AMP
-
-
3'-AMP
-
-
3-Aminopyridine adenine dinucleotide phosphate
-
competitive vs. NADP(H)
4-Chloro-7-nitrobenzo-2-oxa-1,3-diazole
-
34% residual activity at 1 mM
5'-adenosine diphosphate ribose
-
bacteriorhodopsin co-reconstituted enzyme, 29.6% inhibition of thio-NADP+ reduction by NADH in the dark, 13.2% in the light
5'-AMP
5'-[p-(fluorosulfonyl)benzoyl]-adenosine
-
structural analog of adenosine, 2 mM, almost complete inactivation after 25 min, acetylpyridine adenine dinucleotide, NADP+, 5'-AMP, 5'-ADP or a mixture of 2'-AMP and 3'-AMP protect from inactivation, NADPH accelerates the inhibition rate, inhibition rate constant increases 50fold by increasing the pH from 6.0 to 8.5
5,5'-dithiobis(2-nitrobenzoate)
acetyl-CoA
-
-
acetyl-dephospho-CoA
-
competitive vs. NAD(H)
Acetylpyridine adenine dinucleotide
adenosine
ADP
-
-
Butane-2,3-dione
cardiolipin
-
noncompetitive vs. NAD+ and NADPH
D2O
-
-
Dansyl chloride
-
0.25 mM, almost complete inactivation after 8 min, NADP+ or NADPH accelerate inhibition rate
dephospho-CoA
-
competitive vs. NAD(H)
Dicyclohexylcarbodiimide
-
complete inhibition if 0.5 mol are bound to 1 mol of enzyme
diethyldicarbonate
-
inhibition is approx. 50% accelerated in the presence of NAD(H)
Ethoxyformic anhydride
-
2 mM, almost complete inactivation after 6 min, NADP+ or NADPH accelerate inhibition rate
fluorosulfonyl-para-benzyladenosine
-
complete inhibition if 0.5 mol are bound to 1 mol of enzyme
formamide disulfide dihydrochloride
-
0.2 mM, 43% inhibition
glutathione
glutathione disulfide
-
strong, time dependent inhibition of thio-NADP+ reduction by NADH and acetylpyridine adenine dinucleotide reduction by NADPH, 50% inhibition after 40 min incubation in 26.7 mM glutathione disulfide, presence of NADPH accelerates inhibition 20fold
K+
-
200 mM, 50% inhibition at pH 7.9, 300 mM, 40% inhibition at pH 5.5, 95% at pH 8.5
La3+
-
0.1 mM, 50% inhibition at pH 7.0, maximal inhibition at pH 8.0
methylmethane thiosulfonate
N,N'-dicyclohexylcarbodiimide
-
0.3 mM significantly inhibits the the energy-linked NADH-NADP+ reactions, but not the nonenergy-linked NADH-NADP+ transhydrogenation
N,N'-Dicylclohexylcarbodiimide
N-(4-Azido-2-nitrophenyl)-2-aminoethylsulfonate
-
trivial name NAP-taurine, time-dependent inactivation of reconstituted enzyme after photolysis in NAP-taurine loaded vesicles, acetylpyridine adenine dinucleotide stimulates inactivation
N-(Ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline
N-ethylmaleimide
Na+
-
200 mM, 50% inhibition at pH 7.9, 300 mM, 40% inhibition at pH 5.5, 95% at pH 8.5
NADP+
NADPH
p-chloromercuribenzoate
p-Chlororomercuriphenyl sulfonate
-
-
palmitoyl CoA
-
specifically interferes with Nnt activity by competition with NADPH-binding, 20% residual activity at 1 mM
palmitoyl-CoA
Pentane-2,4-dione
-
inactivation of chromatophore complex, 156 mM, approx. 85% inactivation after 30 min, NADPH and NADP+ partially protect, half-maximal protection with 0.015 mM NADPH and 0.030 mM NADP+ respetively
Phenylarsine oxide
Phospholipase A
-
74% inhibition of activity in submitochondrial particles
-
Phospholipase C
-
10-20% inhibition of activity in submitochondrial particles
-
pyridoxal 5'-phosphate
-
0.8 mM, almost complete inactivation after 5 min, 0.4 mM NADP+ or NADPH protect from inactivation, inhibition can be reversed to a considerable extent by L-lysine
reduced acetylpyridine adenine dinucleotide
rotenone
-
with rotenone addition, the NADH-NADP+ activity is inhibited significantly and the remaining activity reflects the nonenergy-linked reaction
S-7-nitrobenzofuran-4-yl-3'-dephospho-CoA
-
strong inhibitor, competitive vs. NAD+ and NADPH
S-7-Nitrobenzofuran-4-yl-CoA
-
strong inhibitor, competitive vs. NADPH, non-competitive vs. NAD+
S-nitrosoglutathione
-
100% inhibition at 3 mM or higher concentration
Sr2+
-
25 mM, 50% inhibition at pH 7.0
Tl+
-
20 mM, 50% inhibition at pH 7.9
triiodothyronine
additional information
-
NNT expression is 2.8fold downregulated in the vastus lateralis muscle of calorie restricted rhesus monkeys
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
asolectin
-
10fold stimulation of partially purified enzyme
-
Carbonyl cyanide m-chlorophenylhydrazone
cardiolipin
-
10fold stimulation of partially purified enzyme with Escherichia coli cardiolipin
H2O2
-
136% increase of activity at 0.5 mM
lecithin
-
5fold stimulation of partially purified enzyme
light
-
forward reaction in chromatophores is accelerated more than 20fold during illumination with photosynthetically active light
-
Lipids
-
-
-
lysolecithin
-
aprrox. 0.3%, 150% activation of activity in chromatophore extracts
lysophosphatidylcholine
-
-
phosphatidylcholine
-
-
Phospholipids
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.028 - 0.166
Acetylpyridine adenine dinucleotide
0.028 - 0.125
NAD+
0.0017 - 0.066
NADH
0.0017 - 0.04
NADP+
0.0051 - 0.12
NADPH
2.6 - 3.4
NMNH
0.02 - 0.8
oxidized acetylpyridine adenine dinucleotide
-
0.008 - 0.063
thio-NADP+
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02 - 1.44
oxidized acetylpyridine adenine dinucleotide
-
0.0667 - 81.7
reduced acetylpyridine adenine dinucleotide
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.59 - 8
2'-AMP
0.003
2,4-Dinitrophenyl-3'-dephospho-CoA
-
-
0.4
3'-5'-AMP
-
-
0.7
3'-AMP
-
-
0.1
3-aminopyridine dinucleotide phosphate
-
approximate value
-
0.3 - 3.7
5'-AMP
0.2
acetyl-CoA
-
-
0.011
acetyl-dephospho-CoA
-
-
0.007 - 0.116
Acetylpyridine adenine dinucleotide
0.5
adenosine
-
-
0.3
ADP
-
-
0.2
CoA
0.009
dephospho-CoA
-
-
2.5
Mg2+
-
at pH 7.0
0.17
NADP+
-
vs. NADPH
0.0035 - 0.26
NADPH
0.00015 - 0.01
palmitoyl-CoA
0.093 - 0.12
reduced acetylpyridine adenine dinucleotide
0.0003
S-7-nitrobenzofuran-4-yl-3'-dephospho-CoA
-
-
0.0026
S-7-Nitrobenzofuran-4-yl-CoA
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.044
-
activity in membranes of strain AB1450
0.1
-
membrane bound mutant enzyme with a direct linker between alpha and beta subunits; purified mutant enzyme with a direct linker between alpha and beta subunits, forward reaction
0.19
-
reduction of acetylpyridine adenine dinucleotide by NADPH
0.2
-
reduction of NADP+ by NADH driven by electron transport, cysteine-free enzyme reconstituted in membrane vesicles
0.26
-
activity in inside-out membrane vesicles at pH 7.4, presence of an uncoupler i.e. carbonylcyanide-m-chlorophylhydrazone results in 2fold stimulation
0.3
-
membrane bound mutant enzyme with a 18 residues long linker between alpha and beta subunits
0.4
-
purified mutant enzyme with a 18 residues long linker between alpha and beta subunits, forward reaction
0.42
-
reduction of NADP+ by NADH driven by electron transport, wild-type enzyme reconstituted in membrane vesicles
0.6
-
membrane bound mutant enzyme with a 32 residues long linker between alpha and beta subunits
0.7
-
purified mutant enzyme with a 32 residues long linker between alpha and beta subunits, forward reaction
0.8
-
purified mutant enzyme with a direct linker between alpha and beta subunits, reverse reaction
0.9
-
purified enzyme, forward reaction
1.4
-
membrane bound enzyme, reverse reaction
1.9
-
reduction of acetylpyridine adenine dinucleotide by NADPH, cysteine-free enzyme reconstituted in membrane vesicles
2.6
-
partially purified enzyme
3
-
reduction of acetylpyridine adenine dinucleotide by NADPH, wild-type enzyme reconstituted in membrane vesicles
3.9
-
purified mutant enzyme with a 18 residues long linker between alpha and beta subunits, reverse reaction
5.5
-
purified mutant enzyme with a 32 residues long linker between alpha and beta subunits, reverse reaction
9.6
-
purified enzyme, reverse reaction
12 - 15
-
-
13.6
-
partially purified enzyme, assay in the presence of Escherichia coli phospholipids
16.2
-
-
22
-
partially purified enzyme from strain W6
24.6
-
reduction of 3-acetylpyridine adenine dinucleotide
29.9
-
purified enzyme from strain JM83
35.6
-
reduction of 3-acetylpyridine adenine dinucleotide
42
-
purified mutant enzyme with a 32 residues long linker between alpha and beta subunits, cyclic reaction
46
-
purified mutant enzyme with a 18 residues long linker between alpha and beta subunits, cyclic reaction
50.8
-
native enzyme with acetylpyridine-NAD+ a n NADPH as substrates, pH 7.0
62.3
-
-
63
-
purified enzyme, cyclic reaction
additional information
-
no activity detected with the insertion mutants
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.5
-
rapid decline above, cyclic reaction
6
-
reverse reaction catalyzed by H91E mutant enzyme, 20% of wild-type enzyme activity
6.2 - 6.3
-
-
7 - 8
-
reduction of oxidized acetylpyridine adenine dinucleotide by NADPH in inside-out membrane vesicles
7
-
reduction of NAD+
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7 - 6.7
-
less than 50% of maximal activity above and below
5.5 - 8.5
-
-
additional information
-
titration of Escherichia coli transhydrogenase domain III with bound NADP+ or NADPH studied by NMR reveals no pH-dependent conformational change in the physiological pH range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
high expression level
Manually annotated by BRENDA team
-
very strong expression
Manually annotated by BRENDA team
-
bone marrow-derived, Nnt mRNA and protein are expressed in resting macrophages and down-regulated in response to inflammatory stimuli such as lipopolysaccharide. Nnt mRNA and protein expression are further repressed in fully activated macrophages; bone marrow-derived, Nnt mRNA and protein are expressed in resting macrophages and down-regulated in response to inflammatory stimuli such as lipopolysaccharide. Nnt mRNA and protein expression are repressed in fully activated macrophages
Manually annotated by BRENDA team
-
high expression level
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Rhodospirillum rubrum (strain ATCC 11170 / ATH 1.1.1 / DSM 467 / LMG 4362 / NCIB 8255 / S1)
Sinorhizobium meliloti (strain SM11)
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)
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)
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
6784
-
x * 6784, soluble component, amino acid analysis
28000
-
dIII fragement, SDS-PAGE
40000
-
dI fragement, SDS-PAGE
47000
-
alpha2,beta2, 2 * 50000 + 2 * 47000
48000
-
x * 53000 + x * 48000, SDS-PAGE
48667
-
alpha2,beta2, 2 * 53906 + 2 * 48667, calculation from nucleotide sequence
50000
-
alpha2,beta2, 2 * 50000 + 2 * 47000
53000
-
x * 53000 + x * 48000, SDS-PAGE
53906
-
alpha2,beta2, 2 * 53906 + 2 * 48667, calculation from nucleotide sequence
54000
-
x * 54000, immunoblot with antibodies against beef heart enzyme
66000
-
2 * 66000, SDS-PAGE
97000
-
x * 97000, SDS-PAGE
109065
-
2 * 109065, monomer is composed of three domains: a 430 residue long N-terminal hydrophilic domain called dI, a 400 residue long central hydrophobic domain that intercalates into the membrane called dII, and a 200 residue long C-terminal hydrophilic domain called dIII
109212
-
2 * 109212, calculated from cDNA sequence
110000
111500
-
2 * 111500, SDS-PAGE
115000
-
2 * 115000, SDS-PAGE
120000
155000
196000
-
-
206000 - 249000
-
cross-linking of purified enzyme with 10.9 mM dimethyl suberimidate dihydrochloride
210000 - 230000
250000
-
-
278000
-
radiation inactivation, hydrodynamic properties
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
solution of isolated dIII domains
tetramer
trimer
additional information
-
the protein has three components: dI binds NADH, dIII binds NADP+, and dII spans the membrane. Transhydrogenase is a dimer of two dI-dII-dIII monomers. The two catalytic sites alternate during turnover
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospholipoprotein
-
5 mol loosely bound phospholipids, 9 mol tightly bound phospholipids
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
dIII domain
-
dimers of dI domains using hanging-drop vapor diffusion method
-
hanging-drop method, crystal structures of the NAD(H)-binding domain I of transhydrogenase in the absence as well as in the presence of oxidized and reduced substrate. The structures are determined at 1.9-2.0 A resolution
dIII domain
-
dIII domain in its thio-NADP+ form
-
dIII domain in the presence of NADPH
-
(dI.Q132N)2dIII trimer
-
crystal structure of domain I i.e. alpha1 subunit, with and without bound NADH, 1.8 A resolution without NADH, 1.9 A with NADH bound
-
crystal structure of domainI/domain III complex
-
dI2dIII complex in its thio-NAD+/NADP+ form
-
dI2dIII complex with bound NAD+ and NADP+, NADH and NADPH, and ADP-ribose and NADPH
-
dI2dIII1 complex and dI domain
-
dI2dIII1 complex with bound NAD+ and 1,4,5,6-tetrahydronicotinamide adenine dinucleotide phosphate the dI2dIII1 complex with bound ,4,5,6-tetrahydronicotinamide adenine dinucleotide and NADP+. dI is the NAD(H)-binding component of transhydrogenase. dIII is the NADP(H)-binding component
-
dIII domain, vapor diffusion method
domain dIII, domain dI dimer and dI2dIII complex in the presence of limiting NADH using the vapor diffusion method
-
solution structure solved by NMR
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6
-
activity decreases as the pH approaches 6.0, although a second minor peak is evident at pH 5.0, the activity of the nonenergy-linked reaction decreases with medium acidification and activity loss is most notable at pH 4.0
695903
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
5 min, soluble component, complete inactivation
44
-
2 min, solubilized membrane component, 50% inactivation
45
-
incubation of depleted membranes, inactivation of membrane transhydrogenase component
48
-
2 min, membrane particles, 50% inactivation
50
-
complete protection in the presence of NADPH, half-maximal protection with 0.01 mM NADPH
53
-
incubation for 3 min, 10 mM Mg2+ almost completely protect from inactivation
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
cations prevent from tryptic inactivation, 95% protection with Mn2+, 89% with Ca2+ and 79% with Mg2+
-
fusion protein is substantially more stable than wild type enzyme upon storage at 4°C
-
inactivation during prolonged column chromatography
-
purified enzyme is inactivated at 4°C even in presence of dithiothreitol
-
solubilized membrane component is unstable loosing all activity when stored overnight at 4°C or -70°C, stable at -70°C in the presence of 0.025 mM NADP+ for several months, inactivation by refreezing after thawing
-
trypsinolysis is stimulated several fold by NADPH and NADP+, half-maximal stimulation with 0.001-0.002 mM NADPH and 0.002-0.003 mM NADP+, Mg2+ protects from NADP+ stimulated inactivation
-
unstable, loses 30-50% activity within 48 h at 4°C, remainder of the enzyme becomes inactive after 2 weeks, complete inactivation after freezing at -20°C or -70°C overnight, enzyme is very susceptible to trypsinolysis, NADH protects from tryptic inactivation, NADPH promotes tryptic inactivation
-
urea, 6 M, inactivation
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1,1-dimethylbutanol
-
inactivation
Acetone
-
used for conversion of mitochondria to an acetone powder causes inactivation
Ethanol
-
10%, 40°C, inactivation
n-Butanol
-
inactivation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15°C, 20 mM sodium tricine buffer, pH 7.6, 1 mM dithiothreitol, 0.2% Triton X-100, 30% glycerol, 4 weeks, no loss of activity
-
-70°C or 4°C, Rhodospirillum rubrum membrane component, inactivation in 1 day, stabilization by NADP+
-
-80°C, 25% glycerol
-
4°C, 0.1 M sodium phosphate buffer, pH 7.5, 1 mM dithiothreitol, 0.05% sodium cholate
-
4°C, 50 mM Tris-HCl, pH 7.8, 1 mM, EDTA, 1 mM, dithiothreitol, 1 week, 10% loss of activity
-
4°C, fusion protein is substantially more stable than wild type enzyme upon storage
-
4°C, reconstituted with phospholipids, stable for at least 2 months
-
4°C, unsoluble component: 0.1 M glycyl-glycine buffer, pH 8.0, 10% sucrose, unstable, soluble factor: 120 h with 0.015 mM dithiothreitol stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
affinity chromatography on NADP+/NAD+-agarose gels, partial purification
-
affinity chromatography; NaCl wash, Triton X-100 extraction, affinity chromatography on immobilized NAD+
-
FPLC in the presence of 0.05 or 0.1% Triton X-100, comparison of methods
-
fractionation of submitochondrial particles, DEAE-Sepharose, hydroxyapatite
-
fusion protein using His-tag and on calmodulin-sepharose
-
His-tagged H91E mutant enzyme
-
immunoexclusion chromatography
-