title: Reactivity and chemical synthesis of L-pyrrolysine- the 22(nd) genetically encoded amino acid.
abstract: L-pyrrolysine, the 22(nd) genetically encoded amino acid, was previously deduced to be (4R, 5R)-4-substituted-pyrroline-5-carboxylate attached to the epsilon-nitrogen of lysine based on the crystal structure of the M. barkeri monomethylamine methyltransferase (MtmB). To confirm L-pyrrolysine's identity, structures of MtmB have been determined following treatment with hydroxylamine, N-methylhydroxylamine, or dithionite. Analysis of these structures has provided additional support for the presence of the pyrroline ring and, together with previous mass spectroscopy data, has led us to assign the C(4)-substituent to a methyl group. Based on this assignment, synthetic L-pyrrolysine was prepared by chemical methods. Detailed study of this chemically synthesized L-pyrrolysine has allowed us to characterize its physical properties, to study its chemical stability, and to elucidate the role of its C(4) substituent. Future applications of this synthetic L-pyrrolysine include its in vivo incorporation into recombinant proteins.
authors.affiliation: Department of Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Hao
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
authors.forename: Patrick T
authors.initials: PT
authors.lastname: Kang
authors.forename: Jitesh A
authors.initials: JA
authors.lastname: Soares
authors.forename: Tsuneo K
authors.initials: TK
authors.lastname: Ferguson
authors.forename: Judith
authors.initials: J
authors.lastname: Gallucci
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Krzycki
authors.forename: Michael K
authors.initials: MK
authors.lastname: Chan
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins; D018360:Crystallography, X-Ray; D004227:Dithionite / Q000737:chemistry; D019811:Hydroxylamine / Q000737:chemistry; D006898:Hydroxylamines / Q000737:chemistry; D008239:Lysine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry*; D017021:Methanosarcina barkeri / Q000201:enzymology*; D008780:Methyltransferases / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure
chemical_list: D019843:Archaeal Proteins; D006898:Hydroxylamines; D004227:Dithionite; D019811:Hydroxylamine; D008780:Methyltransferases; C113200:monomethylamine methyltransferase; C456839:pyrrolysine; D008239:Lysine; C018338:N-methylhydroxylamine