Literature summary for 1.4.9.1 extracted from

Cavalieri, C.; Biermann, N.; Vlasie, M.D.; Einsle, O.; Merli, A.; Ferrari, D.; Rossi, G.L.; Ubbink, M.
Structural comparison of crystal and solution states of the 138 kDa complex of methylamine dehydrogenase and amicyanin from Paracoccus versutus (2008), Biochemistry, 47, 6560-6570.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli	Paracoccus versutus
expressed in Escherichia coli BL21 cells	Paracoccus versutus

Crystallization (Commentary)

Crystallization (Comment)	Organism
MADH-amicyanin binary complex, hanging drop vapour diffusion method, in 28-29.5% PEG8000, 0.2 M Li2SO4, and 0.1 M phosphate (pH 6.5)	Paracoccus versutus
the crystal structure of the complex of MADH and amicyanin is determined to 2.5 A using the hanging-drop method. Enzyme is a heterotetramer consisting of two heavy chains and two light chains. The heavy chain of MADH folds into a characteristic, seven-blade beta-propeller domain and contains an N-terminal extension (residues 1-80) that wraps around the neighboring light chain, fixing it to the tetrameric enzyme. The light subunit consists mainly of loop regions with only four beta-strands, stabilized by a total of six disulfide bridges, containing the active site of the enzyme, a tryptophan tryptophylquinone moiety formed by Trp57 covalently linked to Trp108	Paracoccus versutus

Crystallization (Comment)

Organism

MADH-amicyanin binary complex, hanging drop vapour diffusion method, in 28-29.5% PEG8000, 0.2 M Li2SO4, and 0.1 M phosphate (pH 6.5)

Paracoccus versutus

the crystal structure of the complex of MADH and amicyanin is determined to 2.5 A using the hanging-drop method. Enzyme is a heterotetramer consisting of two heavy chains and two light chains. The heavy chain of MADH folds into a characteristic, seven-blade beta-propeller domain and contains an N-terminal extension (residues 1-80) that wraps around the neighboring light chain, fixing it to the tetrameric enzyme. The light subunit consists mainly of loop regions with only four beta-strands, stabilized by a total of six disulfide bridges, containing the active site of the enzyme, a tryptophan tryptophylquinone moiety formed by Trp57 covalently linked to Trp108

Paracoccus versutus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
KCl	the rate of transfer of the first electron from methylamine-reduced MADH to amicyanin is increased by monovalent cations	Paracoccus versutus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
14200	-	SDS-PAGE, beta subunit	Paracoccus versutus
43300	-	SDS-PAGE, alpha subunit	Paracoccus versutus
138000	-	complex of methylamine dehydrogenase and amicyanin, gel filtration	Paracoccus versutus

Organism

Organism	UniProt	Comment	Textmining
Paracoccus versutus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
DEAE column chromatography	Paracoccus versutus
using a DEAE column	Paracoccus versutus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
dithionite + amicyanin + H2O	two-electron-reduced MADH is obtained by exposing the enzyme either to a 3fold molar excess of methylamine or to 2 mM dithionite	Paracoccus versutus	?	-	?
methylamine + acceptor + H2O	-	Paracoccus versutus	formaldehyde + NH3 + reduced acceptor	-	?
methylamine + amicyanin + H2O	two-electron-reduced MADH is obtained by exposing the enzyme either to a 3fold molar excess of methylamine or to 2 mM dithionite. The complex of MADH and amicyanin in solution is studied using nuclear magnetic resonance. Signals of perdeuterated, 15N-enriched amicyanin bound to MADH are observed. Chemical shift perturbation analysis indicates that the dissociation rate constant is 250/sec and that amicyanin assumes a well-defined position in the complex in solution. The most affected residues are in the interface observed in the crystal structure, whereas smaller chemical shift changes extend to deep inside the protein	Paracoccus versutus	formaldehyde + reduced amicyanin + NH3	-	?
phenylethylamine + acceptor + H2O	-	Paracoccus versutus	phenylacetaldehyde + NH3 + reduced acceptor	-	?

Subunits

Subunits	Comment	Organism
heterotetramer	2 * 43300 + 2 * 14200, X-ray crystallography	Paracoccus versutus
heterotetramer	crystal structure, enzyme shows a alpha2beta2 structure, consisting of two heavy chains and two light chains, alpha subunit: 43300 Da, beta subunit: 14200 Da	Paracoccus versutus

Synonyms

Synonyms	Comment	Organism
MADH	-	Paracoccus versutus
methylamine dehydrogenase	-	Paracoccus versutus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.77	-	amicyanin	methylamine-reduced MADH, pH 7.5 using a Ca-HEPES puffer	Paracoccus versutus
3.1	-	amicyanin	dithionite-reduced MADH, pH 7.5 using a Na,K-phosphate buffer and 0.2 M KCl	Paracoccus versutus
74	-	amicyanin	methylamine-reduced MADH, pH 7.5 using a Na,K-phosphate buffer and 0.2 M KCl	Paracoccus versutus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Paracoccus versutus

Cofactor

Cofactor	Comment	Organism	Structure
heme c	-	Paracoccus versutus
tryptophan tryptophylquinone	-	Paracoccus versutus