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methylamine + H2O + amicyanin
formaldehyde + NH3 + reduced amicyanin
-
-
-
?
1,6-diaminohexane + acceptor + H2O
?
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
1,7-diaminoheptane + acceptor + H2O
?
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
1-aminopentane + acceptor + H2O
pentanal + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
2-phenylethylamine + acceptor + H2O
2-phenylacetaldehyde + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
benzylamine + acceptor + H2O
benzaldehyde + NH3 + reduced acceptor
benzylamine + H2O + ferricyanide
benzaldehyde + NH3 + reduced ferricyanide
-
-
-
-
?
butylamine + acceptor + H2O
butanal + NH3 + reduced acceptor
ethylamine + acceptor + H2O
acetaldehyde + NH3 + reduced acceptor
histamine + acceptor + H2O
? + NH3 + reduced acceptor
-
-
-
-
?
methylamine + acceptor + H2O
formaldehyde + NH3 + reduced acceptor
methylamine + acceptor + H2O
methanal + NH3 + reduced acceptor
methylamine + amicyanin + H2O
formaldehyde + NH3 + reduced amicyanin
-
-
-
-
?
methylamine + H2O + 2 amicyanin
formaldehyde + NH3 + 2 reduced amicyanin
methylamine + H2O + 2,6-dichloroindophenol
formaldehyde + NH3 + reduced 2,6-dichloroindophenol
-
-
-
-
r
methylamine + H2O + 2,6-dichloroindophenol + phenazine ethosulfate
formaldehyde + NH3 + reduced phenazine ethosulfate + ?
-
-
-
-
r
methylamine + H2O + amicyanin
formaldehyde + ammonia + reduced amicyanin
methylamine + H2O + K3Fe(CN)6
formaldehyde + NH3 + reduced K3Fe(CN)6
-
-
-
-
r
n-butylamine + H2O + ferricyanide
butanal + NH3 + reduced ferricyanide
-
-
-
-
?
phenylethylamine + acceptor + H2O
phenylacetaldehyde + NH3 + reduced acceptor
-
-
-
-
?
propylamine + acceptor + H2O
propionaldehyde + NH3 + reduced acceptor
RCH2NH2 + acceptor + H2O
RCHO + NH3 + reduced acceptor
tryptamine + acceptor + H2O
?
-
-
-
-
?
additional information
?
-
benzylamine + acceptor + H2O
benzaldehyde + NH3 + reduced acceptor
-
-
-
-
?
benzylamine + acceptor + H2O
benzaldehyde + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
butylamine + acceptor + H2O
butanal + NH3 + reduced acceptor
-
-
-
-
?
butylamine + acceptor + H2O
butanal + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
ethylamine + acceptor + H2O
acetaldehyde + NH3 + reduced acceptor
-
-
-
-
?
ethylamine + acceptor + H2O
acetaldehyde + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
methylamine + acceptor + H2O
formaldehyde + NH3 + reduced acceptor
-
-
-
-
?
methylamine + acceptor + H2O
formaldehyde + NH3 + reduced acceptor
-
acceptor: amicyanin
-
-
?
methylamine + acceptor + H2O
formaldehyde + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
methylamine + acceptor + H2O
methanal + NH3 + reduced acceptor
-
-
-
-
?
methylamine + acceptor + H2O
methanal + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol or amicyanin
-
-
?
methylamine + H2O + 2 amicyanin
formaldehyde + NH3 + 2 reduced amicyanin
-
-
-
?
methylamine + H2O + 2 amicyanin
formaldehyde + NH3 + 2 reduced amicyanin
-
-
-
?
methylamine + H2O + 2 amicyanin
formaldehyde + NH3 + 2 reduced amicyanin
-
-
-
-
?
methylamine + H2O + amicyanin
formaldehyde + ammonia + reduced amicyanin
-
electron transfer from MADH to cytochrome c-551i does not involve a ternary complex but occurs via a ping-pong mechanism in which amicyanin uses the same interface for the reactions with MADH and cytochrome c-551i. Amicyanin binds tightly to MADH with an interface that matches the one observed in the crystal structure and that mostly overlaps with the binding site for cytochrome c-551i. Amicyanin can react rapidly with cytochrome c-551i, but association of amicyanin with MADH inhibits this reaction
-
-
?
methylamine + H2O + amicyanin
formaldehyde + ammonia + reduced amicyanin
-
P96A and P96G mutations in amycyanin do not affect the spectroscopic or redox properties of amicyanin but increase the Kd value for complex formation with MADH and alter the kinetic mechanism for the interprotein elcetron transfer reaction. The crystal structure of P96G amicyanin is very similar to that of native amicyanin, but in addition to the change in Pro96, the side chains of residues Phe97 and Arg99, which make contacts with MADH that are important for stabilizing the amicyanin-MADH complex, are oriented differently
-
-
?
propylamine + acceptor + H2O
propionaldehyde + NH3 + reduced acceptor
-
-
-
-
?
propylamine + acceptor + H2O
propionaldehyde + NH3 + reduced acceptor
-
acceptor: phenazine ethosulfate/2,6-dichlorophenolindophenol
-
-
?
RCH2NH2 + acceptor + H2O
RCHO + NH3 + reduced acceptor
-
acceptor 2,6-dichlorophenolindophenol
-
-
?
RCH2NH2 + acceptor + H2O
RCHO + NH3 + reduced acceptor
-
acceptor phenazine ethosulfate or amicyanin
-
-
?
RCH2NH2 + acceptor + H2O
RCHO + NH3 + reduced acceptor
-
acceptor potassium ferricyanide, phenazine ethosulfate, 2,6-dichlorophenolindophenol
-
-
?
additional information
?
-
-
amicyanin ami catalyzes the electron transfer from MADH to the terminal oxidase, without the need for any c-type cytochrome. In the absence of either MADH or cytochrome aa3, amicanin is not capable of oxygen reduction on the same time scale. The oxygen consumption depends nearly linearly on the amicyanin concentration up to at least 100 microM. Experiments demonstrate a remarkable number of possibilities for the electron transfer. The interactions appear to be governed exclusively by the electrostatic nature of each of the proteins. Paracoccus denitrificans provides a pool of cytochromes for efficient electron transfer via weak, ill-defined interactions
-
-
?
additional information
?
-
methylamine dehydrogenase (MADH) requires the cofactor tryptophan tryptophylquinone (TTQ) for activity
-
-
?
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tryptophan tryptophylquinone
-
cysteine tryptophylquinone
quinoid cofactor
-
alpha subunit contains unknown quinoid cofactor
-
tryptophan tryptophylquinone
cysteine tryptophylquinone
-
-
cysteine tryptophylquinone
-
cofactor is derived from a pair of gene-encoded amino acids by post-translational modification
heme c
-
contains 1 mol heme c per mol enzyme in the alpha subunit
heme c
-
two heme c cofactors mediate the transfer of the substrate-derived electrons from cysteine tryptophylquinone to an external electron acceptor
tryptophan tryptophylquinone
-
-
tryptophan tryptophylquinone
-
tryptophan tryptophylquinone
-
contains a tryptophan tryptophylquinone prosthetic group
tryptophan tryptophylquinone
-
each beta-subunit possesses a prosthetic group
tryptophan tryptophylquinone
-
a two-electron redox cofactor, each beta subunit of the heterotetrameric enzyme NADH possesses a tryptophan tryptophylquinone, TTQ, protein-derived cofactor. TTQ is formed by post-translational modification of two tryptophan residues of the preMADH polypeptide chain through the diheme enzyme MauG. This six-electron oxidation of preMADH requires long-range electron transfer as the structure of the MauG-preMADH complex reveals that the shortest distance between the modified residues of preMADH and the nearest heme of MauG is 14.0 A, overview
tryptophan tryptophylquinone
-
MauG is a diheme enzyme that catalyzes the final steps in the biosynthesis of the cofactor tryptophan tryptophylquinone in the enzyme
tryptophan tryptophylquinone
TTQ
tryptophan tryptophylquinone
TTQ, methylamine dehydrogenase requires the cofactor tryptophan tryptophylquinone for activity. TTQ is a posttranslational modification that results from an 8-electron oxidation of two specific tryptophans in the MADH beta-subunit, betaTrp57 and betaTrp108. The final 6-electron oxidation is catalyzed by the unusual c-type di-heme enzyme, MauG. The di-ferric enzyme can react with H2O2, but atypically for c-type hemes the di-ferrous enzyme can react with O2 as well. In both cases, an unprecedented bis-Fe(IV) redox state is formed, composed of a ferryl heme (Fe(IV)=O) and the second heme as Fe(IV) stabilized by His-Tyr axial ligation. Bis-Fe(IV) MauG acts as a potent 2-electron oxidant. Catalysis is long-range and requires a hole hopping electron transfer mechanism. TTQ structure analysis, overview
tryptophan tryptophylquinone
TTQ, the catalytic cofactor of enzyme MADH. Activator enzyme MauG is involved in TTQ biosynthesis. Mutation of Trp93 of MauG to tyrosine causes loss of bound Ca2+ and alters the kinetic mechanism of tryptophan tryptophylquinone cofactor biosynthesis. The substrate for MauG-dependent TTQ biosynthesis is preMADH
tryptophan tryptophylquinone
TTQ, the catalytic cofactor of enzyme MADH. It is not an exogenous cofactor but is instead derived from posttranslational modifications of the beta subunits of MADH
tryptophan tryptophylquinone
TTQ, the catalytic cofactor of enzyme MADH. It is not an exogenous cofactor but is instead derived from posttranslational modifications of the beta subunits of MADH as evidenced from the crystal structure of MADH. Kinetic mechanism of MauG-dependent TTQ biosynthesis, overview
tryptophan tryptophylquinone
TTQ, the quinone cofactor of the enzyme. QhpG is involved in the quinone cofactor formation
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0.021 - 0.72
1,6-diaminohexane
0.007 - 0.38
1,7-Diaminoheptane
0.004 - 2.5
1-aminopentane
0.12
2,6-dichloroindophenol
-
-
0.014
phenazine ethosulfate
-
-
0.021
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
0.031
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme betaI107N
0.099
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
0.17
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.72
1,6-diaminohexane
-
pH 7.5, 30°C, wild-type enzyme
0.007
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
0.057
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
0.068
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme betaI107N
0.29
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.38
1,7-Diaminoheptane
-
pH 7.5, 30°C, wild-type enzyme
0.004
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme betaI107N
0.047
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
0.13
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.25
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
2.5
1-aminopentane
-
pH 7.5, 30°C, wild-type enzyme
0.007
Butylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
0.088
Butylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.24
Butylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
0.37
Butylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
0.87
Butylamine
-
pH 7.5, 30°C, wild-type enzyme
0.019
ethylamine
-
pH 7.5, 30°C, wild-type enzyme
0.34
ethylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.36
ethylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
0.84
ethylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
9.2
ethylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
0.004
methylamine
-
pH 8.5, 30°C, wild-type enzyme, acceptor amicyanin
0.005
methylamine
-
pH 7.5, 30°C, wild-type enzyme, acceptor amicyanin
0.0059
methylamine
-
with amicyanin as electron acceptor
0.0064
methylamine
-
with phenylazine as electron acceptor
0.009
methylamine
-
pH 7.5, 30°C, wild-type enzyme
0.015
methylamine
-
pH 7.5, 30°C, wild-type enzyme, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
0.021
methylamine
-
pH 8.5, 30°C, wild-type enzyme, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
0.06
methylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
0.069
methylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.25
methylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
12.2
methylamine
-
pH 8.5, 30°C, mutant enzyme betaD32N, acceptor amicyanin
14.9
methylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
16.7
methylamine
-
pH 7.5, 30°C, mutant enzyme betaD32N, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
19.6
methylamine
-
pH 7.5, 30°C, mutant enzyme betaD32N, acceptor amicyanin
22.4
methylamine
-
pH 8.5, 30°C, mutant enzyme betaD32N, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
0.006
Propylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
0.008
Propylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
0.036
Propylamine
-
pH 7.5, 30°C, wild-type enzyme
0.2
Propylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
1.3
Propylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
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3.4 - 43
1,6-diaminohexane
3.4 - 32
1,7-Diaminoheptane
1.5
2,6-dichloroindophenol
-
-
3.1
2-Phenylethylamine
-
pH 7.5, 30°C
7.1
phenazine ethosulfate
-
-
additional information
additional information
-
turnover numbers for deuterated substrates
-
3.4
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
16
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme betaI107N
17
1,6-diaminohexane
-
pH 7.5, 30°C, wild-type enzyme
26
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme betaI107V
43
1,6-diaminohexane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
3.4
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
15
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme betaI107N
19
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme betaI107V
27
1,7-Diaminoheptane
-
pH 7.5, 30°C, wild-type enzyme
32
1,7-Diaminoheptane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
3.4
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme betaI107V
4.2
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme betaI107N
5.4
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme alphaF55I
17
1-aminopentane
-
pH 7.5, 30°C, wild-type enzyme
20
1-aminopentane
-
pH 7.5, 30°C, mutant enzyme alphaF55A
3.8
benzylamine
-
pH 7.5, 30°C
4.2
Butylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
5.4
Butylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
5.8
Butylamine
-
pH 7.5, 30°C
14
Butylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
22
Butylamine
-
pH 7.5, 30°C, wild-type enzyme
34
Butylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
4.5
ethylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
6.2
ethylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
15
ethylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
23
ethylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
24
ethylamine
-
pH 7.5, 30°C, wild-type enzyme
0.14
methylamine
-
pH 7.5, 30°C, mutant enzyme betaD32N, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
0.19
methylamine
-
pH 8.5, 30°C, mutant enzyme betaD32N, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
0.47
methylamine
-
pH 7.5, 30°C, mutant enzyme betaD32N, acceptor amicyanin
1.2
methylamine
-
pH 8.5, 30°C, mutant enzyme betaD32N, acceptor amicyanin
2
methylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
13.6
methylamine
-
with phenylazine as electron acceptor
18
methylamine
-
pH 8.5, 30°C, wild-type enzyme, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
20
methylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
23
methylamine
-
pH 7.5, 30°C, wild-type enzyme, acceptor amicyanin
26
methylamine
-
pH 8.5, 30°C, wild-type enzyme, acceptor amicyanin
30
methylamine
-
pH 7.5, 30°C, wild-type enzyme
34
methylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
35
methylamine
-
pH 7.5, 30°C, wild-type enzyme, acceptor phenazine ethosulfate/2,6-dichlorophenolindophenol
48.5
methylamine
-
with amicyanin as electron acceptor
77
methylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
3.1
Propylamine
-
pH 7.5, 30°C, mutant enzyme betaI107V
4.1
Propylamine
-
pH 7.5, 30°C, mutant enzyme betaI107N
5.2
Propylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55I
24
Propylamine
-
pH 7.5, 30°C, mutant enzyme alphaF55A
27
Propylamine
-
pH 7.5, 30°C, wild-type enzyme
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evolution
the qhp genes are very widely distributed, not only in many Gram-negative species but also in a few Gram-positive bacteria, bacterial distribution of qhp and associated genes, overview. The subunits constituting QHNDH are encoded by ORF1 (alpha subunit), ORF4 (beta subunit), and ORF3 (gamma subunit). Of the other genes in the operon, ORF2 encodes an [Fe-S] cluster and S-adenosylmethionine (SAM)-binding protein, a member of the radical SAM superfamily, and ORF5 encodes a protein of approximately 22.5 kDa belonging to subfamily S8A of peptidase family S8 (the subtilisin family) with the conserved Asp/His/Ser catalytic triad characteristic of this subfamily
malfunction
the genes mauF and mauE are membrane proteins with no homology to characterized proteins, and are thought to be involved in transport of MADH subunits into the periplasm. Knocking out either gene leads to no detectable beta-subunit in the periplasm, and an unusual beta-subunit leader sequence is consistent with it being trafficked by a specific transporter. The loss of mauF and mauE additionally leads to a drastic reduction in alpha-subunit. The third gene, mauD, is homologous to disulfide isomerases, and is likely specific to the MADH beta-subunit, which has six disulfides. In the absence of mauD, periplasmic alpha-subunit levels are close to normal, but again there is no detectable beta-subunit implying that the disulfides are key to beta-subunit stability. When the final required gene, mauG, is knocked out, there are normal levels of MADH alpha- and beta-subunit in the periplasm, but no methylamine dehydrogenase activity is present. This has focused attention on the mauG gene product as a likely participant in TTQ biosynthesis
physiological function
MADH catalyzes the oxidative deamination of methylamine to formaldehyde and ammonia, a reaction which allows the host bacterium to use methylamine as a sole source of carbon, nitrogen and energy. MADH donates the electrons which it extracts from methylamine to the mauC gene product, a type 1 copper protein named amicyanin, which in turn transfers electrons to cytochrome c-551i. The catalytic cofactor of MADH is tryptophan tryptophyquinone, TTQ
physiological function
the diheme enzyme MauG catalyzes a six electron oxidation that is required for the posttranslational modification of a precursor of methylamine dehydrogenase (preMADH) to complete the biosynthesis of its protein-derived cofactor, tryptophan tryptophylquinone (TTQ). The substrate for MauG that undergoes this posttranslational modification is a precursor protein of MADH (preMADH). It possesses a monohydroxylated residue betaTrp57. The reactions catalyzed by MauG occur in the following order: covalent cross-linking of monohydroxylated betaTrp57 to betaTrp108, incorporation of a second oxygen atom into the side chain of betaTrp57, and oxidation of the quinol species to the quinone. Catalysis requires long-range electron transfer because preMADH does not make direct contact with either heme of MauG. The electron transfer occurs via a hole-hopping mechanism in which Trp residues of MauG are reversibly oxidized
physiological function
the diheme enzyme MauG catalyzes oxidative post-translational modifications of a protein substrate, precursor protein of methylamine dehydrogenase (preMADH), that binds to the surface of MauG. Tinding of preMADH to MauG affects both the coordination state of the ferric high-spin heme, and the kinetic mechanism of the autoreduction of the bis-FeIV hemes. Binding sructure analysis, overview
additional information
-
MauG is a diheme enzyme responsible for the posttranslational modification of two tryptophan residues in pre-MADH to form the tryptophan tryptophylquinone, TTQ, cofactor of methylamine dehydrogenase. MauG catalyzes a six electron oxidation to complete TTQ biosynthesis. Oxidizing equivalents may be provided by three mol of either O2, plus an electron donor, or H2O2
additional information
-
steady-state MauG-depedent TTQ biosynthesis using quinol MADH as a substrate and single-turnover kinetics of the reaction of bis-Fe(IV) MauG with quinol MADH, overview
additional information
QM/MM molecular dynamics simulations at room temperature generate a multidimensional thermal free-energy landscape without restriction of the degrees of freedom beyond a multidimensional reaction subspace mapping two rather similar pathways for the underlying proton transfer to one of two aspartate carboxyl oxygen atoms, termed OD1 and OD2, which hydrogen bond with Thr122 and Trp108, respectively. Despite significant large-amplitude motion perpendicular to the one-dimensional proton transfer coordinate, due to fluctuations of the donor-acceptor distance of about 3 a, it is found that the one-dimensional proton transfer free-energy profiles are essentially identical to the minimum free-energy pathways on the multidimensional free-energy landscapes for both proton transfer channels. Proton transfer to one of the acceptor oxygen atoms (the OD2 site) is slightly favored in methylamine dehydrogenase both kinetically and thermodynamically. Modeling is based on the crysta structure of the substrate-free enzyme MADH from Paracoccus denitrificans resolved at 1.75 A, PDB ID 2BBK
additional information
structural features of the gamma subunit clearly indicate that it must undergo multiple posttranslational modifications before it can form an active QHNDH complex with the alpha and beta subunits. The qhpG gene encodes a putative FAD-dependent monooxygenase, which is required for the generation of the quinone cofactor in the gamma subunit. The qhpR gene encodes an AraC family transcriptional regulator, which activates expression of the qhp operon in response to the addition of n-butylamine to the culture medium. The structural genes encoding the three QHNDH subunits constitute an operon harboring six apparent open reading frames (ORFs) that are transcribed in a coordinated manner upon addition of amines to the culture medium. QhpF serves as an efflux ABC transporter for translocation of the gamma subunit of QHNDH into the periplasm
additional information
-
structural features of the gamma subunit clearly indicate that it must undergo multiple posttranslational modifications before it can form an active QHNDH complex with the alpha and beta subunits. The qhpG gene encodes a putative FAD-dependent monooxygenase, which is required for the generation of the quinone cofactor in the gamma subunit. The qhpR gene encodes an AraC family transcriptional regulator, which activates expression of the qhp operon in response to the addition of n-butylamine to the culture medium. The structural genes encoding the three QHNDH subunits constitute an operon harboring six apparent open reading frames (ORFs) that are transcribed in a coordinated manner upon addition of amines to the culture medium. QhpF serves as an efflux ABC transporter for translocation of the gamma subunit of QHNDH into the periplasm
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betaD32N
-
preparation contains a major species with six disulfides but no oxygen incorporated into betaTrp57 and a minor species with both oxygens incorporated, which is active. 1000fold increase in KM-value for methylamine
betaD76N
-
mutant enzyme is completely inactive
F55A
-
mutation of the alpha subunit
F55E
-
mutation of the alpha subunit
T122A
-
the presence of Thr122 has a deleterious effect on the proton transfer step that is proposed to determine the rate of the reaction, the substitution of Thr122 by Ala does not significantly modify the preference of the proton by atom OD2 of Asp76
alphaF55A
-
1656fold increase in Km-value for methylamine compared to wild-type enzyme, 484fold increase in Km-value for ethylamine compared to wild-type enzyme, 36fold increase in Km-value for propylamine compared to wild-type enzyme, 3.6fold decrease in Km-value for butylamine compared to wild-type enzyme, 53.2fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 34.3fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 54.3fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
alphaF55A
-
inactivation by the mechanism-based inhibitor cyclopropylamine is accompanied by the formation of a covalent cross-link between the alpha and beta subunits of the enzyme. No cross-linking is seen with mutant enzymes alphaF55A or alphaF55I mutant enzymes
alphaF55A
-
mutation decreases the affinity for binding of monovalent cations, Na+ or K+. 1656fold increase in Km-value for methylamine compared to wild-type enzyme, 484fold increase in Km-value for ethylamine compared to wild-type enzyme, 36fold increase in Km-value for propylamine compared to wild-type enzyme, 3.6fold decrease in Km-value for butylamine compared to wild-type enzyme, 53.2fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 34.3fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 54.3fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
alphaF55A
-
mutation increases the rate of the electron transfer reaction from the fully reduced tryptophan tryptophylquinone tryptophan tryptophylquinone methylamine dehydrogenase to amicyanin. Little difference in the overal structure of alphaF55A in complex with its electron acceptors, amicyanin and cytochrome c-551i, relative to the native complex. There are significant changes in the solvent content of the active site and substrate channel
alphaF55I
-
1.2fold decrease in Km-value for methylamine compared to wild-type enzyme, 18.9fold increase in Km-value for ethylamine compared to wild-type enzyme, 5.6fold increase in Km-value for propylamine compared to wild-type enzyme, 4.2fold decrease in Km-value for butylamine compared to wild-type enzyme, 10fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 7.3fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 6.7fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzym. Ability to discriminate between amines of different chain length is abolished
alphaF55I
-
inactivation by the mechanism-based inhibitor cyclopropylamine is accompanied by the formation of a covalent cross-link between the alpha and beta subunits of the enzyme. No cross-linking is seen with mutant enzymes alphaF55A or alphaF55I mutant enzymes
alphaF55I
-
mutation has no effect on binding of monovalent cation. 1.2fold decrease in Km-value for methylamine compared to wild-type enzyme, 18.9fold increase in Km-value for ethylamine compared to wild-type enzyme, 5.6fold increase in Km-value for propylamine compared to wild-type enzyme, 4.2fold decrease in Km-value for butylamine compared to wild-type enzyme, 10fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 7.3fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 6.7fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
betaI107N
-
27.8fold increase in Km-value for methylamine compared to wild-type enzyme, 44.2fold increase in Km-value for ethylamine compared to wild-type enzyme, 8.5fold decrease in Km-value for propylamine compared to wild-type enzyme, 124fold decrease in Km-value for butylamine compared to wild-type enzyme, 62.5fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 23.2fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 5.6fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
betaI107N
-
27.8fold increase in Km-value for methylamine compared to wild-type enzyme, 44.2fold increase in Km-value for ethylamine compared to wild-type enzyme, 8.5fold decrease in Km-value for propylamine compared to wild-type enzyme, 124fold decrease in Km-value for butylamine compared to wild-type enzyme, 62.5fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 23.2fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 5.6fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme. Mutant enzyme exhibity a strong preference for 1-aminopentane compared to strong preference for methylamine of the wild-type enzyme
betaI107V
-
7.7fold increase in Km-value for methylamine compared to wild-type enzyme, 17.9fold increase in Km-value for ethylamine compared to wild-type enzyme, 6fold decrease in Km-value for propylamine compared to wild-type enzyme, 9.9fold decrease in Km-value for butylamine compared to wild-type enzyme, 19.2fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 4.2fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 1.3fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
betaI107V
-
mutant enzyme exhibits a strong preference for propylamine compared to strong preference for methylamine of the wild-type enzyme. 7.7fold increase in Km-value for methylamine compared to wild-type enzyme, 17.9fold increase in Km-value for ethylamine compared to wild-type enzyme, 6fold decrease in Km-value for propylamine compared to wild-type enzyme, 9.9fold decrease in Km-value for butylamine compared to wild-type enzyme, 19.2fold decrease in Km-value for 1-aminopentane compared to wild-type enzyme, 4.2fold decrease in Km-value for 1-6-diaminohexane compared to wild-type enzyme, 1.3fold decrease in Km-value for 1,7-diaminoheptane compared to wild-type enzyme
additional information
generation of of the gene disrupted mutant strains PdDELTAqhpF, PdDELTAqhpG, and PdDELTAqhpR
additional information
-
generation of of the gene disrupted mutant strains PdDELTAqhpF, PdDELTAqhpG, and PdDELTAqhpR
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Brooks, H.B.; Jones, L.H.; Davidson, V.L.
Deuterium kinetic isotope effect and stopped-flow kinetic studies of the quinoprotein methylamine dehydrogenase
Biochemistry
32
2725-2729
1993
Paracoccus denitrificans
brenda
Takagi, K.; Torimura, M.; Kawaguchi, K.; Kano, K.; Ikeda, T.
Biochemical and electrochemical characterization of quinohemoprotein amine dehydrogenase from Paracoccus denitrificans
Biochemistry
38
6935-6942
1999
Paracoccus denitrificans, Paracoccus denitrificans IFO 12442
brenda
Sun, D.; Davidson, V.L.
Re-engineering monovalent cation binding sites of methylamine dehydrogenase: effects on spectral properties and gated electron transfer
Biochemistry
40
12285-12291
2001
Paracoccus denitrificans
brenda
Bao, L.; Sun, D.; Tachikawa, H.; Davidson, V.L.
Improved sensitivity of a histamine sensor using an engineered methylamine dehydrogenase
Anal. Chem.
74
1144-1148
2002
Paracoccus denitrificans
brenda
Datta, S.; Ikeda, T.; Kano, K.; Mathews, F.S.
Structure of the phenylhydrazine adduct of the quinohemoprotein amine dehydrogenase from Paracoccus denitrificans at 1.7 A resolution
Acta Crystallogr. Sect. D
59
1551-1556
2003
Paracoccus denitrificans
brenda
Sun, D.; Chen, Z.W.; Mathews, F.S.; Davidson, V.L.
Mutation of alphaPhe55 of methylamine dehydrogenase alters the reorganization energy and electronic coupling for its electron transfer reaction with amicyanin
Biochemistry
41
13926-13933
2002
Paracoccus denitrificans
brenda
Sun, D.; Ono, K.; Okajima, T.; Tanizawa, K.; Uchida, M.; Yamamoto, Y.; Mathews, F.S.; Davidson, V.L.
Chemical and kinetic reaction mechanisms of quinohemoprotein amine dehydrogenase from Paracoccus denitrificans
Biochemistry
42
10896-10903
2003
Paracoccus denitrificans
brenda
Davidson, V.L.
Probing mechanisms of catalysis and electron transfer by methylamine dehydrogenase by site-directed mutagenesis of alpha Phe55
Biochim. Biophys. Acta
1647
230-233
2003
Paracoccus denitrificans
brenda
Sun, D.; Davidson, V.L.
Inter-subunit cross-linking of methylamine dehydrogenase by cyclopropylamine requires residue alphaPhe55
FEBS Lett.
517
172-174
2002
Paracoccus denitrificans
brenda
Wang, Y.; Sun, D.; Davidson, V.L.
Use of indirect site-directed mutagenesis to alter the substrate specificity of methylamine dehydrogenase
J. Biol. Chem.
277
4119-4122
2002
Paracoccus denitrificans
brenda
Jones, L.H.; Pearson, A.R.; Tang, Y.; Wilmot, C.M.; Davidson, V.L.
Active site aspartate residues are critical for tryptophan tryptophylquinone biogenesis in methylamine dehydrogenase
J. Biol. Chem.
280
17392-17396
2005
Paracoccus denitrificans
brenda
Sun, D.; Li, X.; Mathews, F.S.; Davidson, V.L.
Site-directed mutagenesis of proline 94 to alanine in amicyanin converts a true electron transfer reaction into one that is kinetically coupled
Biochemistry
44
7200-7206
2005
Paracoccus denitrificans
brenda
Ma, J.K.; Carrell, C.J.; Mathews, F.S.; Davidson, V.L.
Site-directed mutagenesis of proline 52 to glycine in amicyanin converts a true electron transfer reaction into one that is conformationally gated
Biochemistry
45
8284-8293
2006
Paracoccus denitrificans
brenda
Wang, Y.; Li, X.; Jones, L.H.; Pearson, A.R.; Wilmot, C.M.; Davidson, V.L.
MauG-dependent in vitro biosynthesis of tryptophan tryptophylquinone in methylamine dehydrogenase
J. Am. Chem. Soc.
127
8258-8259
2005
Paracoccus denitrificans
brenda
Ono, K.; Okajima, T.; Tani, M.; Kuroda, S.; Sun, D.; Davidson, V.L.; Tanizawa, K.
Involvement of a putative [Fe-S]-cluster-binding protein in the biogenesis of quinohemoprotein amine dehydrogenase
J. Biol. Chem.
281
13672-13684
2006
Paracoccus denitrificans
brenda
Pierdominici-Sottile, G.; Echave, J.; Palma, J.
Molecular dynamics study of the active site of methylamine dehydrogenase
J. Phys. Chem. B
110
11592-11599
2006
Paracoccus denitrificans
brenda
Ma, J.K.; Wang, Y.; Carrell, C.J.; Mathews, F.S.; Davidson, V.L.
A single methionine residue dictates the kinetic mechanism of interprotein electron transfer from methylamine dehydrogenase to amicyanin
Biochemistry
46
11137-11146
2007
Paracoccus denitrificans
brenda
Li, X.; Fu, R.; Liu, A.; Davidson, V.L.
Kinetic and physical evidence that the diheme enzyme MauG tightly binds to a biosynthetic precursor of methylamine dehydrogenase with incompletely formed tryptophan tryptophylquinone
Biochemistry
47
2908-2912
2008
Paracoccus denitrificans
brenda
Pierdominici-Sottile, G.; Marti, M.A.; Palma, J.
The role of residue Thr122 of methylamine dehydrogenase on the proton transfer from the iminoquinone intermediate to residue Asp76
Chem. Phys. Lett.
456
243-246
2008
Paracoccus denitrificans
-
brenda
Ranaghan, K.E.; Masgrau, L.; Scrutton, N.S.; Sutcliffe, M.J.; Mulholland, A.J.
Analysis of classical and quantum paths for deprotonation of methylamine by methylamine dehydrogenase
ChemPhysChem
8
1816-1835
2007
Paracoccus denitrificans
brenda
Pearson, A.R.; Pahl, R.; Kovaleva, E.G.; Davidson, V.L.; Wilmot, C.M.
Tracking X-ray-derived redox changes in crystals of a methylamine dehydrogenase/amicyanin complex using single-crystal UV/Vis microspectrophotometry
J. Synchrotron Radiat.
14
92-98
2007
Paracoccus denitrificans
brenda
Fujieda, N.; Mori, M.; Ikeda, T.; Kano, K.
The silent form of quinohemoprotein amine dehydrogenase from Paracoccus denitrificans
Biosci. Biotechnol. Biochem.
73
524-529
2009
Paracoccus denitrificans
brenda
Shin, S.; Abu Tarboush, N.; Davidson, V.L.
Long-range electron transfer reactions between hemes of MauG and different forms of tryptophan tryptophylquinone of methylamine dehydrogenase
Biochemistry
49
5810-5816
2010
Paracoccus denitrificans
brenda
Jensen, L.M.; Sanishvili, R.; Davidson, V.L.; Wilmot, C.M.
In crystallo posttranslational modification within a MauG/pre-methylamine dehydrogenase complex
Science
327
1392-1394
2010
Paracoccus denitrificans
brenda
Choi, M.; Sukumar, N.; Mathews, F.S.; Liu, A.; Davidson, V.L.
Proline 96 of the copper ligand loop of amicyanin regulates electron transfer from methylamine dehydrogenase by positioning other residues at the protein-protein interface
Biochemistry
50
1265-1273
2011
Paracoccus denitrificans
brenda
Meschi, F.; Wiertz, F.; Klauss, L.; Cavalieri, C.; Blok, A.; Ludwig, B.; Heering, H.A.; Merli, A.; Rossi, G.L.; Ubbink, M.
Amicyanin transfers electrons from methylamine dehydrogenase to cytochrome c-551i via a ping-pong mechanism, not a ternary complex
J. Am. Chem. Soc.
132
14537-14545
2010
Paracoccus denitrificans
brenda
Meschi, F.; Wiertz, F.; Klauss, L.; Blok, A.; Ludwig, B.; Merli, A.; Heering, H.A.; Rossi, G.L.; Ubbink, M.
Efficient electron transfer in a protein network lacking specific interactions
J. Am. Chem. Soc.
133
16861-16867
2011
Paracoccus denitrificans
brenda
Sukumar, N.; Choi, M.; Davidson, V.L.
Replacement of the axial copper ligand methionine with lysine in amicyanin converts it to a zinc-binding protein that no longer binds copper
J. Inorg. Biochem.
105
1638-1644
2011
Paracoccus denitrificans
brenda
de la Lande, A.; Babcock, N.S.; Rezac, J.; Sanders, B.C.; Salahub, D.R.
Surface residues dynamically organize water bridges to enhance electron transfer between proteins
Proc. Natl. Acad. Sci. USA
107
11799-11804
2010
Paracoccus denitrificans
brenda
Yukl, E.T.; Jensen, L.M.; Davidson, V.L.; Wilmot, C.M.
Structures of MauG in complex with quinol and quinone MADH
Acta Crystallogr. Sect. F
69
738-743
2013
Paracoccus denitrificans (Q51658)
brenda
Yukl, E.T.; Goblirsch, B.R.; Davidson, V.L.; Wilmot, C.M.
Crystal structures of CO and NO adducts of MauG in complex with pre-methylamine dehydrogenase: implications for the mechanism of dioxygen activation
Biochemistry
50
2931-2938
2011
Paracoccus denitrificans (Q51658)
brenda
Choi, M.; Shin, S.; Davidson, V.L.
Characterization of electron tunneling and hole hopping reactions between different forms of MauG and methylamine dehydrogenase within a natural protein complex
Biochemistry
51
6942-6949
2012
Paracoccus denitrificans
brenda
Abu Tarboush, N.; Jensen, L.M.; Wilmot, C.M.; Davidson, V.L.
A Trp199Glu MauG variant reveals a role for Trp199 interactions with pre-methylamine dehydrogenase during tryptophan tryptophylquinone biosynthesis
FEBS Lett.
587
1736-1741
2013
Paracoccus denitrificans (A1BBA0 and A1BB97)
brenda
Shin, S.; Davidson, V.L.
MauG, a diheme enzyme that catalyzes tryptophan tryptophylquinone biosynthesis by remote catalysis
Arch. Biochem. Biophys.
544
112-118
2014
Paracoccus denitrificans (P22619 AND P29894)
brenda
Shin, S.; Feng, M.; Davidson, V.L.
Mutation of Trp93 of MauG to tyrosine causes loss of bound Ca2+ and alters the kinetic mechanism of tryptophan tryptophylquinone cofactor biosynthesis
Biochem. J.
456
129-137
2013
Paracoccus denitrificans (P22619 AND P29894)
brenda
Shin, S.; Yukl, E.T.; Sehanobish, E.; Wilmot, C.M.; Davidson, V.L.
Site-directed mutagenesis of Gln103 reveals the influence of this residue on the redox properties and stability of MauG
Biochemistry
53
1342-1349
2014
Paracoccus denitrificans (P22619 AND P29894)
brenda
Nakai, T.; Deguchi, T.; Frebort, I.; Tanizawa, K.; Okajima, T.
Identification of genes essential for the biogenesis of quinohemoprotein amine dehydrogenase
Biochemistry
53
895-907
2014
Paracoccus denitrificans (P22619 AND P29894), Paracoccus denitrificans
brenda
Zelleke, T.; Marx, D.
Free-energy landscape and proton transfer pathways in oxidative deamination by methylamine dehydrogenase
Chemphyschem
18
208-222
2017
Paracoccus denitrificans (P22619 AND P29894)
brenda
Wilmot, C.; Yukl, E.
MauG A di-heme enzyme required for methylamine dehydrogenase maturation
Dalton Trans.
42
3127-3135
2013
Paracoccus denitrificans (P22619 AND P29894)
-
brenda
Feng, M.; Ma, Z.; Crudup, B.F.; Davidson, V.L.
Properties of the high-spin heme of MauG are altered by binding of preMADH at the protein surface 40 A away
FEBS Lett.
591
1566-1572
2017
Paracoccus denitrificans (P22619 AND P29894)
brenda
Jo, M.; Shin, S.; Choi, M.
Intra-electron transfer of amicyanin from newly derived active site to redox potential tuned type 1 copper site
Appl. Biol. Chem.
61
181-187
2018
Paracoccus denitrificans (P22619)
-
brenda
Jeoung, S.; Shin, S.; Choi, M.
Copper-binding energetics of amicyanin in different folding states
Metallomics
12
273-279
2020
Paracoccus denitrificans (P22619), Paracoccus denitrificans
brenda