Changes in Protein Patterns of Pea Plants Systemically Infected with Red Clover Mottle Virus

Marcinka, K; Röhring, C; Kluge, Siegfried

doi:10.1016/s0015-3796(11)80006-5

Cited by 11 publications

(6 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…SDS-gels were used to determine the molecular mass of subunits of the hydroxylase and to screen for purity. The SDS-gels were silver stained (28).…”

Section: (Ii) Reductase (Component C)mentioning

confidence: 99%

Purification and Characterization of the Soluble Methane Monooxygenase of the Type II Methanotrophic Bacterium Methylocystis sp. Strain WI 14

Große

Laramée

Wendlandt

et al. 1999

Appl Environ Microbiol

View full text Add to dashboard Cite

Methane monooxygenase (MMO) catalyzes the oxidation of methane to methanol as the first step of methane degradation. A soluble NAD(P)H-dependent methane monooxygenase (sMMO) from the type II methanotrophic bacterium WI 14 was purified to homogeneity. Sequencing of the 16S rDNA and comparison with that of other known methanotrophic bacteria confirmed that strain WI 14 is very close to the genusMethylocystis. The sMMO is expressed only during growth under copper limitation (<0.1 μM) and with ammonium or nitrate ions as the nitrogen source. The enzyme exhibits a low substrate specificity and is able to oxidize several alkanes and alkenes, cyclic hydrocarbons, aromatics, and halogenic aromatics. It has three components, hydroxylase, reductase and protein B, which is involved in enzyme regulation and increases sMMO activity about 10-fold. The relative molecular masses of the native components were estimated to be 229, 41, and 18 kDa, respectively. The hydroxylase contains three subunits with relative molecular masses of 57, 43, and 23 kDa, which are present in stoichiometric amounts, suggesting that the native protein has an α2β2γ2structure. We detected 3.6 mol of iron per mol of hydroxylase by atomic absorption spectrometry. sMMO is strongly inhibited by Hg2+ions (with a total loss of enzyme activity at 0.01 mM Hg2+) and Cu2+, Zn2+, and Ni2+ ions (95, 80, and 40% loss of activity at 1 mM ions). The complete sMMO gene sequence has been determined. sMMO genes from strain WI 14 are clustered on the chromosome and show a high degree of homology (at both the nucleotide and amino acid levels) to the corresponding genes fromMethylosinus trichosporium OB3b, Methylocystissp. strain M, and Methylococcus capsulatus (Bath).

show abstract

“…SDS-gels were used to determine the molecular mass of subunits of the hydroxylase and to screen for purity. The SDS-gels were silver stained (28).…”

Section: (Ii) Reductase (Component C)mentioning

confidence: 99%

Purification and Characterization of the Soluble Methane Monooxygenase of the Type II Methanotrophic Bacterium Methylocystis sp. Strain WI 14

Große

Laramée

Wendlandt

et al. 1999

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…The reaction was started by addition of appropriate amounts of ethanol and propionaldehyde, respectively. The SDS-gels were silver stained as described by MARCINKA (1992).…”

Section: De/rninmentioning

confidence: 99%

Purification and properties of methanol dehydrogenase from Methylocystis sp. GB 25

Große

Wendlandt

Kleber

1997

J. Basic Microbiol.

View full text Add to dashboard Cite

Methanol dehydrogenase (MDH) from Methylocystis sp. GB 25, which belongs to the group II of methanotrophic bacteria, is able to catalyse the oxidation of methanol to formate directly. The enzyme was purified 20-fold by a 5 step procedure to electrophoretic homogeneity. After cell disruption by French press, about 95% of MDH-activity was found in the soluble fraction. The relative molecular mass of the native enzyme has been estimated to be 122 kDa by gel filtration and 115 kDa by the method of Hedrick and Smith (1968). It seems to be composed of two identical subunits with a relative molecular mass of 62 kDa (estimated by SDS gel electrophoresis). The isoelectric point was found to be about 8.3. The amino terminal sequence shows a strong similarity to the alpha-chain of MDH from the facultative methylotrophic bacterium Methylobacterium extorquens AM1. PQQ, the probable prosthetic group of MDH, could be detected in the supernatant of the culture by using the apoenzyme of a membrane-bound glucose dehydrogenase from Pseudomonas aeruginosa but not absolutely in the absorption spectra of the enzyme after DEAE-chromatography. The purified MDH has an optimum activity at pH 9.0 and at 45 degrees C. MDH of Methylocystis sp. GB 25 oxidises only primary alcohols from methanol to heptanol and aldehydes from formaldehyde to propionaldehyde and the glutaraldehyde, respectively. The estimated Km-values show no dependence upon the chain length of substrates.

show abstract

“…SDS-polyacrylamide gels were used to determine molecular masses of subunits of the proteins and to screen for purity. The SDS-polyacrylamide gels were silver-stained (Marcinka et al 1992).…”

Section: Purification Of CIImentioning

confidence: 99%

Biotransformation of crotonobetaine to L(-)- carnitine in Proteus sp.

Engemann

Elßner

Kleber

2001

Archives of Microbiology

View full text Add to dashboard Cite

Two proteins, component I (CI) and component II (CII), catalyze the biotransformation of crotonobetaine to L(-)-carnitine in Proteus sp. CI was purified to electrophoretic homogeneity from cell-free extracts of Proteus sp. The N-terminal amino acid sequence of CI showed high similarity (80%) to the caiB gene product from Escherichia coli O44K74, which encodes the L(-)-carnitine dehydratase. CI alone was unable to convert crotonobetaine into L(-)-carnitine even in the presence of the cosubstrates crotonobetainyl-CoA or gamma-butyrobetainyl-CoA, which are essential for this biotransformation. The relative molecular mass of CI was determined to be 91.1 kDa. CI is composed of two identical subunits of molecular mass 43.6 kDa. The isoelectric point is 5.0. CII was purified to electrophoretic homogeneity from cell-free extracts of Proteus sp. and its N-terminal amino acid sequence showed high similarity (75%) to the caiD gene product of E. coli O44K74. The relative molecular mass of CII was shown to be 88.0 kDa, and CII is composed of three identical subunits of molecular mass 30.1 kDa. The isoelectric point of CII is 4.9. For the biotransformation of crotonobetaine to L(-)-carnitine, the presence of CI, CII, and a cosubstrate (crotonobetainyl-CoA or gamma-butyrobetainyl-CoA) were shown to be essential.

show abstract

Changes in Protein Patterns of Pea Plants Systemically Infected with Red Clover Mottle Virus

Cited by 11 publications

References 20 publications

Purification and Characterization of the Soluble Methane Monooxygenase of the Type II Methanotrophic Bacterium Methylocystis sp. Strain WI 14

Purification and Characterization of the Soluble Methane Monooxygenase of the Type II Methanotrophic Bacterium Methylocystis sp. Strain WI 14

Purification and properties of methanol dehydrogenase from Methylocystis sp. GB 25

Biotransformation of crotonobetaine to L(-)- carnitine in Proteus sp.

Contact Info

Product

Resources

About