Publications

• Function of genetically encoded pyrrolysine in corrinoid-dependent methylamine methyltransferases.

  Krzycki J.A.

  Methanogenesis from trimethylamine, dimethylamine or monomethylamine is initiated by a series of corrinoid-dependent methyltransferases. The non-homologous genes encoding the full-length methyltransferases each possess an in-frame UAG (amber) codon that does not terminate translation. The amber co ... >> More

  Methanogenesis from trimethylamine, dimethylamine or monomethylamine is initiated by a series of corrinoid-dependent methyltransferases. The non-homologous genes encoding the full-length methyltransferases each possess an in-frame UAG (amber) codon that does not terminate translation. The amber codon is decoded by a dedicated tRNA, and corresponds to the novel amino acid pyrrolysine in one of the methyltransferases, indicating pyrrolysine to be the 22nd genetically encoded amino acid. Pyrrolysine has the structure of lysine with the (epsilon)N in amide linkage with a pyrroline ring. The reactivity of the electrophilic imine bond is the basis for the proposed function of pyrrolysine in activating and optimally orienting methylamine for methyl transfer to the cobalt ion of a cognate corrinoid protein. This reaction is essential for methane formation from methylamines, and may underlie the retention of pyrrolysine in the genetic code of methanogens. << Less

  Curr Opin Chem Biol 8:484-491(2004) [PubMed] [EuropePMC]

  This publication is cited by 2 other entries.

• Clustered genes encoding the methyltransferases of methanogenesis from monomethylamine.

  Burke S.A., Lo S.L., Krzycki J.A.

  Coenzyme M (CoM) is methylated during methanogenesis from monomethyamine in a reaction catalyzed by three proteins. Using monomethylamine, a 52-kDa polypeptide termed monomethylamine methyltransferase (MMAMT) methylates the corrinoid cofactor bound to a second polypeptide, monomethylamine corrinoi ... >> More

  Coenzyme M (CoM) is methylated during methanogenesis from monomethyamine in a reaction catalyzed by three proteins. Using monomethylamine, a 52-kDa polypeptide termed monomethylamine methyltransferase (MMAMT) methylates the corrinoid cofactor bound to a second polypeptide, monomethylamine corrinoid protein (MMCP). Methylated MMCP then serves as a substrate for MT2-A, which methylates CoM. The genes for these proteins are clustered on 6.8 kb of DNA in Methanosarcina barkeri MS. The gene encoding MMCP (mtmC) is located directly upstream of the gene encoding MMAMT (mtmB). The gene encoding MT2-A (mtbA) was found 1.1 kb upstream of mtmC, but no obvious open reading frame was found in the intergenic region between mtbA and mtmC. A single monocistronic transcript was found for mtbA that initiated 76 bp from the translational start. Separate transcripts of 2.4 and 4.7 kb were detected, both of which carried mtmCB. The larger transcript also encoded mtmP, which is homologous to the APC family of cationic amine permeases and may therefore encode a methylamine permease. A single transcriptional start site was found 447 bp upstream of the translational start of mtmC. MtmC possesses the corrinoid binding motif found in corrinoid proteins involved in dimethylsulfide- and methanol-dependent methanogenesis, as well as in methionine synthase. The open reading frame of mtmB was interrupted by a single in-frame, midframe, UAG codon which was also found in mtmB from M. barkeri NIH. A mechanism that circumvents UAG-directed termination of translation must operate during expression of mtmB in this methanogen. << Less

  J. Bacteriol. 180:3432-3440(1998) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Reconstitution of monomethylamine:coenzyme M methyl transfer with a corrinoid protein and two methyltransferases purified from Methanosarcina barkeri.

  Burke S.A., Krzycki J.A.

  Methanogenesis from methylamines requires the intermediate methylation of 2-mercaptoethanesulfonate (CoM). In vitro reconstitution of CoM methylation with monomethylamine was achieved with three purified proteins: a monomethylamine corrinoid protein (MMCP), the "A" isozyme of methylcobamide:CoM me ... >> More

  Methanogenesis from methylamines requires the intermediate methylation of 2-mercaptoethanesulfonate (CoM). In vitro reconstitution of CoM methylation with monomethylamine was achieved with three purified proteins: a monomethylamine corrinoid protein (MMCP), the "A" isozyme of methylcobamide:CoM methyltransferase (MT2-A), and a newly isolated protein termed monomethylamine methyltransferase (MMAMT).MMAMT is a 170-kDa protein with 52-kDa subunits. The MMAMT polypeptide was rate-limiting for methyl transfer until at a 2-fold molar excess over MMCP. MMAMT is a monomethylamine:MMCP methyltransferase, since methylation of MMCP required MMAMT but not MT2-A. MMCP and MMAMT formed a complex detectable by size exclusion high pressure liquid chromatography. Methyl group transfer from methyl-MMCP to CoM was mediated by MT2-A, since methyl iodide:CoM methyl transfer by MMCP and MT2-A did not require MMAMT. MT2-M, an isozyme of MT2-A, was inactive in MMCP-dependent methyl transfer. Immunodepletion of MMCP from the extract inhibited CoM methylation with monomethylamine but not dimethylamine. Purified MMCP reconstituted activity in immunodepleted extracts. These results show that MMCP is the major corrinoid protein for methanogenesis from monomethylamine detectable in extracts and that it interacts with two methyltransferases. MMAMT functions as a MMA:MMCP methyltransferase, while MT2-A functions as a methyl-MMCP:CoM methyltransferase. << Less

  J. Biol. Chem. 272:16570-16577(1997) [PubMed] [EuropePMC]