Involvement of methyltransferase-activating protein and methyltransferase 2 isoenzyme II in methylamine:coenzyme M methyltransferase reactions in Methanosarcina barkeri Fusaro.
The enzyme systems involved in the methyl group transfer from methanol and from tri- and dimethylamine to 2-mercaptoethanesulfonic acid (coenzyme M) were resolved from cell extracts of Methanosarcina barkeri Fusaro grown on methanol and trimethylamine, respectively. Resolution was accomplished by ammonium sulfate fractionation, anion-exchange chromatography, and fast protein liquid chromatography. The methyl group transfer reactions from tri- and dimethylamine, as well as the monomethylamine:coenzyme M methyltransferase reaction, were strictly dependent on catalytic amounts of ATP and on a protein present in the 65% ammonium sulfate supernatant. The latter could be replaced by methyltransferase-activating protein isolated from methanol-grown cells of the organism. In addition, the tri- and dimethylamine:coenzyme M methyltransferase reactions required the presence of a methylcobalamin:coenzyme M methyltransferase (MT2), which is different from the analogous enzyme from methanol-grown M. barkeri. In this work, it is shown that the various methylamine:coenzyme M methyltransfer steps proceed in a fashion which is mechanistically similar to the methanol:coenzyme M methyl transfer, yet with the participation of specific corrinoid enzymes and a specific MT2 isoenzyme.
Wassenaar
R W
RW
Department of Microbiology, Faculty of Science, University of Nijmegen, The Netherlands. roelw@sci.kun.nl.
Daas
P J
PJ
Geerts
W J
WJ
Keltjens
J T
JT
van der Drift
C
C
D000255:Adenosine Triphosphate / Q000378:metabolism; D019843:Archaeal Proteins; D004123:Dimethylamines / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism*; D015080:Mesna / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000201:enzymology*; D008744:Methylamines / Q000378:metabolism*; D008780:Methyltransferases / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*
D019843:Archaeal Proteins; D004123:Dimethylamines; D007527:Isoenzymes; D008744:Methylamines; D000255:Adenosine Triphosphate; C034516:dimethylamine; D008780:Methyltransferases; C103170:dimethylamine-coenzyme M methyltransferase; C023894:methylcobalamin-coenzyme M methyltransferase; C103169:mttB protein, Methanosarcina barkeri; D011494:Protein Kinases; C101667:methyltransferase activation protein, Methanosarcina barkeri; C023336:trimethylamine; D015080:Mesna; D000432:Methanol