Identification of genes involved in the acetamidino group modification of the flagellin N-Linked glycan of Methanococcus maripaludis

Gareth M. Jones, John Wu, Yan Ding, Kaoru Uchid, Shin Ichi Aizaw, Anna Robotham, Susan M. Logan, John Kelly, Ken F. Jarrell

研究成果: 雜誌貢獻文章

24 引文 斯高帕斯(Scopus)

摘要

N-linked glycosylation of protein is a posttranslational modification found in all three domains of life. The flagellin proteins of the archaeon Methanococcus maripaludis are known to be modified with an N-linked tetrasaccharide consisting of N-acetylgalactosamine (GalNAc), a diacetylated glucuronic acid (GlcNAc3NAc), an acetylated and acetamidino-modified mannuronic acid with a substituted threonine group (ManNAc3NAmA6Thr), and a novel terminal sugar residue [(5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose]. To identify genes involved in biosynthesis of the component sugars of this glycan, three genes, mmp1081, mmp1082, and mmp1083, were targeted for in-frame deletion, based on their annotation and proximity to glycosyltransferase genes known to be involved in assembly of the glycan. Mutants carrying a deletion in any of these three genes remained flagellated and motile. A strain with a deletion of mmp1081 had lower-molecular-mass flagellins in Western blots. Mass spectrometry of purified flagella revealed a truncated glycan with the terminal sugar absent and the threonine residue and the acetamidino group missing from the third sugar. No glycan modification was seen in either the Δmmp1082 or Δmmp1083 mutant grown in complex Balch III medium. However, a glycan identical to the Δmmp1081 glycan was observed when the Δmmp1082 or Δmmp1083 mutant was grown under ammonia-limited conditions. We hypothesize that MMP1082 generates ammonia and tunnels it through MMP1083 to MMP1081, which acts as the amidotransferase, modifying the third sugar residue of the M. maripaludis glycan with the acetamidino group.
原文英語
頁(從 - 到)2693-2702
頁數10
期刊Journal of Bacteriology
194
發行號10
DOIs
出版狀態已發佈 - 五月 1 2012
對外發佈Yes

    指紋

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

引用此

Jones, G. M., Wu, J., Ding, Y., Uchid, K., Aizaw, S. I., Robotham, A., Logan, S. M., Kelly, J., & Jarrell, K. F. (2012). Identification of genes involved in the acetamidino group modification of the flagellin N-Linked glycan of Methanococcus maripaludis. Journal of Bacteriology, 194(10), 2693-2702. https://doi.org/10.1128/JB.06686-11